Cape Bio Unit 1 Notes
Cape Bio Unit 1 Notes
1.1: Discuss how the structure and properties of water relate to the role that water plays as a medium of
life.
Your body is comprised of numerous elements, which make also combine to form molecules. These
include macronutrients such as carbohydrates (such as starch and glucose, required for release of ATP),
proteins (which are used for growth and repair of cells and also to form hormones) and fats (used for as an
energy store).
However, the molecule that comprises the majority of the human body (more than 70% of a cell’s mass)
is WATER.
First, observe the molecular structure of water. Water consists of two hydrogen atoms COVALENTLY
bonded to one oxygen atom. This means that electrons are shared between them.
On the diagram, you will observe the symbol δ (delta), and a symbol for +ve or –ve charge. In this case,
the OXYGEN has the negative charge and the HYDROGEN atoms have the positive charge.
Water itself is electrically balanced or NEUTRAL. However, there is uneven distribution of these charges
in the structure. This is called a DIPOLE. This allows weak electrical attraction between the water
molecules, which results in COHESION and the ability to undego MASS FLOW. They also result in
HYDROGEN BONDS, which are essential for many biological molecules.
Temp. regulation Its high specific heat capacity and ability to evaporate easily.
‘Universal’ solvent Its tiny charges attract other molecules or ions to form bonds.
Allows mass flow Its H-bonds produce cohesion and surface tension. Suitable for excretion.
Carbohydrates are organic molecules that comprise a ratio of carbon, hydrogen and oxygen. They are
comprised of at least one sugar unit. However, they can be linked together to form increasingly complex
molecules.
MONOSACCHARIDES are the simplest carbohydrate and cannot be further hydrolysed. They are
written with the general formula (CH O) . The ‘n’ depends on the type of sugar. For example, a hexose
2 n
A pentose such as ribose has a value of ‘5’ and is written as C H O . However, modifications occur, such
5 10 5
As previously mentioned, glucose is a HEXOSE, which means it has a six-membered ring consisting of
five carbons and one oxygen. Observe the straight-chain and ring structures of glucose below.
It can be observed that the 6th carbon atom in the ring structure does not exist as part of the ring structure.
As a result of this, glucose tends to alternate between its ring and its chain form. This is why there are two
different types of glucose (alpha and beta).
As previously mentioned, a DISACCHARIDE forms when two monosaccharide molecules are bonded.
When this linkage occurs, it is known as a GLYCOSIDIC bond. These types of bonds are very strong.
One common example of a disaccharide is SUCROSE, which we commonly know as the sugar that is
sweet (such as in sugar cane).
So, what two monosaccharides combine to form sucrose? That would be an ALPHA GLUCOSE and a
BETA FRUCTOSE. What is notable about sucrose is that when it undergoes enzyme breakdown, sucrose
yields two glucose molecules. However, one of those molecules has been reformed from fructose.
In O’ levels you would’ve learned that Benedict’s solution can be used to test for reducing sugar.
However, the addition of HYDROCHLORIC ACID and then SODIUM HYDROXIDE was needed for
non-reducing sugars.
This is because disaccharides such as sucrose have a glycosidic bond that prevents Benedict’s reageant
from reacting with it. The HCl is needed to break that glycosidic bond and the NaOH is needed to
neutralize the HCl.
1.4: Discuss how the molecular structure of starch, glycogen and cellulose relate to their functions in
living organisms.
A polysaccharide can contain thousands of sugar molecules and can be quite large and complex. As a
result, they are insoluble. Not all of them are arranged in long chains, however. Some of them form
compact spirals. Polysaccharide nutrients such as starch must be hydrolysed before they can be absorbed
through the small intestine and into the bloodstream.
GLYCOGEN Energy reserve in Easier to break down into glucose. Usually found in the
animals. LIVER and in MUSCLES.
CELLULOSE Found in cell walls. Always has a straight structure. Very strong due to
Used for structural thousands of hydrogen bonds. Large bundles of them are
support. called FIBRES. Difficult for animals to digest.
The table below will provide a summary of all of this complex information.
Overall shape Linear and spiral Linear, spiral, branches Only linear
• Breaking a covalent bond is called a HYDROLYSIS REACTION, while formation of the bond is
called a CONDENSATION REACTION.
• Hydrolysis reactions use a water molecule during the breakdown of polymers into monomers.
Condensation reactions release a molecule during the formation of a bond. If that molecule is
water, this is known as a DEHYDRATION reaction.
• Examples of dehydration reactions include the formation of SUCROSE (from glucose & fructose)
and the formation of a DIPEPTIDE molecule from two amino acids.
• Hydrogen bonds form between water molecules. HYDROXYL groups (-OH) form hydrogen
bonds because hydrogen is slightly +ve and oxygen is slightly –ve. Dipole or polar molecules are
hydrophilic while non-polar molecules (without dipoles) are hydrophobic.
1.5: Describe the molecular structure of a triglyceride and its role as a source of energy.
Lipids have a similar chemical structure to carbohydrates. The main difference is that they contain a
much higher proportion of HYDROGEN. They also tend to be insoluble in water. The main lipids that
you would have previously learned of are fats and oils, which are used as energy reserves in the body and
also used to provide insulation for organs.
Fats are broken down by the enzyme LIPASE (secreted by the pancreas). This results in the formation of
FATTY ACIDS AND GLYCEROL. These fatty acids can be classified as either saturated or
unsaturated (more on this later).
A TRIGLYCERIDE is comprised of three fatty acids attached to a glycerol molecule. They are insoluble
in water and are HYDROPHOBIC, meaning that they are not attracted to water. The fatty acids contain a
–COOH, which is called a CARBOXYL group. These carboxyl groups react with the –OH groups of
glycerol. This forms a very strong covalent bond called an ESTER BOND.
Thus, think of the glycerol as the ‘backbone’ of the triglyceride structure. Observe the detailed structure
of a glycerol molecule and a triglyceride below:
You should also get familiar with how it is represented in simpler diagrams:
Triglycerides are an energy reserve and are stored in tissues in humans called ADIPOSE tissue.
Accumulation of excess adipose tissue will eventually lead to OBESITY. Studies of fat are constantly
yielding new information and show that fats act almost like endocrine organs, affecting hormonal
secretion and metabolism.
The cells shown are called ADIPOCYTES. ‘White fat’ cells have a much higher concentration of
triglycerides than ‘brown fat’ cells.
Brown fat cells tend to have a high concentration of mitochondria, which regularly ‘burn’ off the energy
reserves.
As previously stated, fatty acids are typically classified into two types: saturated and unsaturated. What
is the main difference between these two?
• A SATURATED fat molecule has its last carbon atom bonded to three hydrogens. Thus, it has
been ‘saturated’ with hydrogen.
This is usually referred to as the ‘bad’ fat, as it forms a dense structure that can contribute to the
build-up of LDL (low-density lipoprotein) cholesterol, leading to coronary heart disease.
• An UNSATURATED fat molecule has at least one carbon atom double-bonded to another,
reducing the amount of hydrogen that is holds.
Observe below to see that it causes a slight bend in the linear structure. Imagine that this bend
prevents the fat from packing too tightly and contributing to arterial plaque build-up.
1.6: Describe the structure of phospholipids and their role in membrane structure and function.
WHAT IS A PHOSPHOLIPID?
Observe the diagram shown. It shows a phospholipid bilayer (which forms the plasma membrane).
Imagine a triglyceride where one of its fatty acids has been replaced by a PHOSPHATE group.
On the diagram, you’ll notice that the phosphate ‘heads’ are HYDROPHILIC while the ‘fatty acid’ tails
are HYDROPHOBIC. If you recall, hydrophobic means they are not attracted to water molecules.
Hydrophilic means they are attracted. So in water, they form this ‘bilayer’ structure. Without this
structure, cells would not be able to keep their organelles together.
1.7: Describe the generalised structure of an amino acid, & the formation & breakage of a peptide bond.
You will recall from O’ Level Biology that proteins are mainly used for cellular growth and repair in the
body. They also form a entire roster of other molecules in the body, including enzymes and hormones. An
amino acid is a single unit and many of these combine to form a protein, just like with monosaccharides
and polysaccharides.
Observe the structure. There is a central carbon atom connected to FOUR other groups. These include:
Amino acids can bond with each other during condensation reactions. The linkages formed are very
strong covalent bonds called PEPTIDE BONDS.
When this occurs, a H atom joins with an –OH to form a water molecule.
WHAT IS A POLYPEPTIDE?
Protein synthesis occurs in the RIBOSOMES of the cells. As previously said, condensation reactions
occur when amino acids are bonded, which produce water molecules.
When many of these amino acids are linked by peptide bonds, the chain itself is called a POLYPEPTIDE.
These polypeptide chains eventually come together to form structures of protein.
The chains can be non-linear in shape. For example, HAEMOGLOBIN (found in the red blood cells) has
four polypeptides connected in a coiled structure.
When polypeptide chains are broken, a water molecule is consumed during a hydrolysis reaction. An
example of this would be when PEPSIN digests proteins in the stomach.
Observe the linkage between two amino acids to form a dipeptide molecule:
WHAT ARE SOME EXAMPLES OF AMINO ACIDS?
There are 20 amino acids. They may be hydrophilic or hydrophobic. Only the ones with side chains (‘R’
groups) that contain ring structures are hydrophobic. Here are a few examples of amino acids.
• Serine – Used in the synthesis of components in the brain cell membranes and neurones.
• Leucine – Involved in increasing lean muscle mass.
• Valine – High levels are associated with insulin resistance and diabetes.
• Tryptophan – Converts to serotonin, which affects mood and sleep
• Aspartic acid – Contributes to the formation of urea.
1.8: Explain the meaning of terms: primary, secondary, tertiary and quaternary structures of proteins.
Recall that proteins are comprised of amino acid units which form polypeptide chains. The way in which
these are sequenced can occur in multiple levels of increasing complexity in proteins, resulting in what
are known as the primary, secondary, tertiary and quaternary structures.
There are four main types of bonds that help form the linkages that hold protein molecules in shape.
Hydrophobic Very weak. Not considered a bond. But Forms between R groups which
Interaction can denature in high heat. contain only C and H atoms.
1.9: Outline the molecular structure of collagen, as an example of a fibrous protein;
Collagen is a protein found in our bodies that is mainly used for STRUCTURAL SUPPORT. It can be
found in areas such as cartilage, bones and tendons. Due to its structural role, its insolubility in water and
its repeating sequences, it is referred to as a FIBROUS protein. This contrasts with GLOBULAR proteins,
such as haemoglobin, antibodies and enzymes, which partake in chemical reactions, are often soluble in
water and the primary structures usually have specific shapes instead of repeated sequences.
As can be seen in the molecular structure, it consists of THREE polypeptide chains. These form three
helical strands, which intertwine and are held together by HYDROGEN bonds.
These collagen molecules form cross-links and form FIBRILS, which form bundles known as FIBRES.
1.10: Carry out tests for reducing and nonreducing sugars, starch, lipids and proteins.
Examples of reducing sugars include GLUCOSE, MALTOSE and FRUCTOSE, while an example of a
non-reducing sugar is SUCROSE. The solution needed to test for both of these is called BENEDICT’S
SOLUTION, a blue liquid that contains copper (II) sulphate.
Upon heating, Cu is reduced to Cu and forms copper (I) oxide in the presence of reducing sugar, which
2+ +
forms a BRICK RED precipitate. Trace amounts of sugars results in a GREEN colour.
FOR NON-REDUCING SUGARS: Recall that sucrose has a GLYCOSIDIC bond. To break this bond,
heat the solution with dilute HCl and then neutralize with SODIUM HYDROXIDE. This will yield
GLUCOSE and FRUCTOSE from the sucrose.
Starch is a polysaccharide that is comprised of amylose and amylopectin. The test for starch presence
involves the addition of IODINE SOLUTION IN POTASSIUM IODIDE (KI/I ). The iodine is able to
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bind to the helical structure of amylose and produce the BLUE-BLACK colour.
Proteins have linkages called PEPTIDE bonds (between the C and N of adjacent amino acids). BIURET
reagent is used to test for proteins, which contains copper (II) sulphate and potassium hydroxide.
When BIURET reagent is added, the copper ions produce a PURPLE colour.
Recall that lipids are hydrophobic and are thus INSOLUBLE in water. To test for the presence of lipids,
ETHANOL is first poured into the sample. The lipid molecules will dissolve in the ethanol. WATER is
then added.
The hydrophobic lipid molecules begin to disassociate from the solution and form an opaque milky white
layer of droplets that float to the top called an EMULSION.
TOPIC 2: CELL STRUCTURE
2.1 and 2.2: Make drawings of typical animal and plant cells as seen under the light microscope and
describe and interpret drawings and electron micrographs of cells;
Colour Image will appear in colour. Image will be in black and white.
Preparation of Living cells and tissues are used. Non- Only non-living and dehydrated cells
specimen living tissues may be used if they are are used. They are cut very thinly and
mounted on a slide in a transparent liquid. placed in a vacuum.
Staining of Cells absorb many different coloured Cells and organelles absorb heavy
specimen stains. metals.
Main advantages - Much more affordable than electron - Much higher resolution
microscopes.
- Much higher magnification
- Slides can last for a very long time.
- Little risk of distortion while viewing.
Recall that light microscopes have a much lower resolution and magnification than electron microscopes.
Photomicrographs therefore are unable to clearly show all of the organelles present in the structures of the
animal and plant cells. When an electron microscope is used to view the structure, the visible image is
called an ULTRASTRUCTURE.
2.4. Compare the structure of typical animal and plant cells;
You also have to be able to look at electron micrographs and label the organelles on the ultrastructure.
The arrangement of these will vary in specialized cells. Look at this mouse’s hepatocyte (liver cell). It is
very dense with membranes from the ER and has many secretory vesicles and lysosomes.
The reason for this being that the liver must form a highly active transport network for proteins, lipids and
sugars. They must also break down many toxins, such as alcohol.
2.5. Describe the structure of a prokaryotic cell. Compare their structure with eukaryotic cells.
Prokaryotes (which means “before the nucleus”) are organisms that have DNA but in a circular or freely
dispersed form not present in a nucleus. They form their own kingdom and includes organisms such as
BACTERIA and ARCHAEA.
Eukaryotes (which means “true nucleus”) have a nucleus and so also have a nuclear envelope and
nucleolus. They also have membrane-bound organelles such as MITOCHONDRIA and
CHLOROPLASTS. They belong to more complex organisms such as animals, plants and protists. So far,
we’ve mainly been looking at eukaryotic cells.
Genetic material No nucleus but contains plasmids Has a nucleus and all internal
(circular DNA) and a nucleoid region of structures. DNA in long strands,
protoplasmic DNA. connected to histones.
Membrane-bound Mitochondria, chloroplasts, Golgi body These same structures are usually
organelles and ER are all absent. present. Chloroplasts only in plants.
Cell wall Made of peptidoglycan (e.g. bacteria). Made of cellulose (in plants)
Flagella Present in many cells (e.g. E. coli Present in a few, such as sperm cells
bacteria). or some protists. Different structure.
Photosynthetic Contains infolds in the plasma membrane Contains chloroplasts, which contain
structures for chlorophyll attachment. chlorophyll.
As previously noted, the word “prokaryote” means “before the nucleus” and are thus this type of
organism is ancient (approximately 3.5 billion years). It was also previously noted that prokaryotes do not
have membrane-bound organelles such as MITOCHONDRIA and CHLOROPLASTS. It was believed a
very long time ago that three main types of cells existed:
It is now thought that the latter two organisms were absorbed by the first, thus giving rise to a new type of
cell that would be able to carry out the processes of PHOTOSYNTHESIS and RESPIRATION. They
were called ENDOSYMBIONTS, which eventually gave rise to other types of organelles and specialized
cells, which led to the rise of many different organisms as time passed.
2.7: Explain the concepts of tissue and organ using as an example the dicotyledonous root and stem.
Cells are known as the basic functional and biological units of all living organisms, whether they are
unicellular or multicellular. Many cells can be SPECIALIZED to perform specific functions, such as red
blood cells containing haemoglobin to carry oxygen, sperm cells having a flagellum or muscle cells being
able to contract.
When multiple cells form groups the carry out the same function, they are known as TISSUES. An
example of this would be multiple cells called neurones forming a tissue called a nerve. Blood is also an
example of a tissue, since it contains many cells, such as red blood cells, lymphocytes and phagocytes.
These tissues are then grouped together to form ORGANS and then ORGAN SYSTEMS. Examples of
organs include the heart, liver, eye, leaf and root.
Observe the structures through a transverse cross-section of a buttercup (Ramunculus) root shown below.
Epidermis Has root hairs to provide large surface area for water absorption.
Cortex/Parenchyma Move water to the centre of the root either through cell walls or cells.
Air spaces Contains oxygen for aerobic respiration. Pathway for rapid diffusion.
Endodermis Waterproof layer to limit capillary action, due to presence of Casparian strips.
Vascular bundle Contains xylem for transporting water (across lignified walls) and phloem for
translocation of sucrose (through phloem sieve elements).
The diagrams above and below depict a transverse section from a stem tissue taken from a Dahlia
specimen. Look at the above plan diagram and label the microscopic image similarly.
Sclerenchyma A very thick, hard layer of tissue used for support. Usually dead cells.
Collenchyma Layer of elongated cells with thick cell walls used for support. Usually alive.
The phosphilipid bilayer forms the basis of the plasma membrane around the cell, separating the inner
cytoplasm from the extracellular content. It is made up of phospholipids, which have HYDROPHILIC
(attracted to water) phosphate heads and HYDROPHOBIC (repelled by water) fatty acid tails.
This creates a double layer with larger proteins and other structures known as the fluid mosaic model. The
model can be thought as a “sea of phospholipids with protein icebergs”.
Membranes are important for transfer of materials, acting as sites for receptors and enzymes. They also
allow passage of electrical signals, such as in the axons of neurones.
Structure Function or Notes
Glycoproteins and Can act as receptor sites to allow binding of certain molecules such as
Glycolipids HORMONES or NEUROTRANSMITTERS.
Extrinsic Proteins Do not penetrate the bilayer. May have glycoproteins attached to them.
Intrinsic Proteins Fixed into structure.. Have hydrophilic and hydrophic regions. The hydrophobic
regions are usually attracted to the lipid tails by HYDROPHOBIC
INTERACTIONS. May also act as ENZYMES.
3.2: Explain the processes of diffusion, facilitated diffusion, osmosis, active transport, endocytosis and
exocytosis.
The plasma membrane allows movement of molecules into and out of the cells. This can happen in a
number of ways. This process can occur without the use of ATP (called PASSIVE transport) or with the
use of ATP (called ACTIVE transport).
These two types of transport usually rely on the creation of a difference in concentrations on both sides
(or a concentration gradient).
Movement of molecules can also occur via VESICLES either from the inside to the exterior of the cell
(EXOCYTOSIS) or from the exterior into the cell (ENDOCYTOSIS).
Diffusion and Facilitated Diffusion are both examples of passive transport, which means that they do not
require the use of ATP.
Using the example with the KMnO crystals, diffusion occurs because the water molecules have an
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They bombard the crystals, causing them to break apart and move outward. This movement will naturally
shift the crystals down a concentration gradient.
We can thus define diffusion as: THE NET MOVEMENT OF MOLECULES OR IONS FROM
REGIONS OF HIGHER TO LOWER CONCENTRATION.
However, the main difference is that DIFFUSION allows molecules to enter the cell by moving through
the phospholipid bilayer (imagine like water draining through a layer of sand). Facilitated diffusion requires
the use of specific pathways called CHANNEL PROTEINS, which form hydrophilic passages. Imagine
these channels like gates that will only allow entry for specific molecules or ions.
1. Unlike the other two, active transport requires the use of ATP.
2. Active transport moves molecules from up or AGAINST a concentration gradient (from a region
of LOW concentration to a region of HIGH concentration).
In exocytosis, a VESICLE is used as the transport sac for the material. The vesicle will move to the
plasma membrane, combine with it and release the contents. This process allows the secretion of
substances such as enzymes and antibodies.
In endocytosis, the substance usually enters the cell through the plasma membrane. Sometimes the cell
changes shape to accommodate the material (such as during PHAGOCYTOSIS in macrophages). The
area becomes enclosed, forms a vesicle and is absorbed by the cytoplasm.
Water molecules are small enough to pass through the tiny spaces in the phosopholipid bilayer, but only
at low rates (due to the hydrophobic fatty acid tails) It thus can be said to be PARTIALLY
PERMEABLE. There are also specialized channels called AQUAPORINS that allow the movement of
water molecules from a higher to lower water potential. Why not say ‘concentration’?
Water potential is depicted as the Greek symbol ‘psi’ (ψ). Think of water potential as “the tendency of
water to leave the solution” or the pressure that will push water molecules across, so the higher the value
is more likely the water molecules will move across the membrane.
• HYPOTONIC solutions have very low conc. of solute and so have a high ψ.
• HYPERTONIC solutions have high conc. of solute and have a low ψ.
You will see water potential usually being represented as a negative (-) number. In fact, the water
potential of pure water at atmospheric pressure (with absolutely no solute in it) has a water potential of
ZERO. The more solute there is, the more negative Ψ becomes, since the solute molecules will attract the
water molecules and restrict their freedom to move.
Recall that about 60% of your body is water. A great amount of that is found in the cells as components of
protoplasm and cytoplasm.
Moisture is also used to line membranes, such as in the alveoli, and water is a main component of blood
plasma. It is an absolute necessity to regulate the water-salt balance in the human body to prevent the
cells from either shrivelling (CRENATION) or bursting (LYSIS).
3.3: Investigate the effects on plant cells of immersion into solutions of different water potentials.
You may recall performing an experiment in O’ Level Biology involving submerging potato cylinders in
solutions of varying sucrose concentrations. You would’ve then compared the final lengths/masses to the
initial lengths/masses of the cylinders to determine whether or not water flowed into the cell or flowed
out.
If a cylinder happens to have no change in length and mass, then it could be assumed that there was no
difference in water potential inside and outside of the cell (ψ = ψ ). The basis of this experiment is to
solution potato
perform trials with multiple sucrose solutions and graph the % change. When there is 0% change, that
would be equal to the water potential of the plant tissue.
Let’s do this sample question below to plot the graph and determine the water potential of the tissue:
Molarity of sucrose sol’n (mol dm ) 0.0 0.1 0.2 0.3 0.4 0.5
-3
% change in mass 24 15 11 2 -4 -8
From the graph, the water potential of the tissue will be found at a molarity of 0.35 mol dm . -3
Solute potential (or ψ ) can be defined as the amount by which a dissolved solute lowers the water
s
potential. Simply put, the higher the solute potential, the lower the water potential. It is represented as a
negative number and the higher the solute potential, the more negative that number is (e.g. 0.50 mol dm -3
If there is too much solute in the cell, water will leave and the cell loses turgor and is said to have
undergone PLASMOLYSIS. There is a point where the cell loses enough internal water pressure that it
stops pressing against the cell wall. As a result, the cell wall stops pushing back. The pressure potential
now becomes zero. This is the moment just before plasmolysis occurs, when the plasma membrane will
begin to retract. This point is called INCIPIENT plasmolysis.
This experiment seeks to determine that point. You can think of it as the point where the water potential
inside is equal to the solute potential inside (ψ = ψ ), that will cause water to start to flow out. To
inside s inside
observe this, the tissue will be observed under a microscope under varying sucrose concentrations.
The higher the sucrose concentration, the more cells will become plasmolysed. However, this will not
happen immediately. There will be a sucrose concentration that will act as a sudden ‘tipping point’. The
aim to determine that point. Observe the sample readings below and plot the graph:
Salt conc. / g dm -3
0.0 0.5 1.3 1.6 1.8 2.1 2.3 2.5 2.7 3.5 4.0 5.0 6.0
% plasmolysis 0 0 0 15 30 60 80 96 100 100 100 100 100
TOPIC 4: ENZYMES
4.1 and 2: Explain that enzymes are globular proteins that catalyse metabolic reactions. Also explain the
mode of action of enzymes in terms of an active site, enzyme and/or substrate complex, lowering of
activation energy and enzyme specificity.
Enzymes are globular proteins. They tend to be involved in metabolic reactions because their TERTIARY
structures (which are folded 3D structures of α helices and β sheets) are quite unique. As a result,
enzymes tend to be involved in specific reactions instead of structural roles, like fibrous proteins (e.g.
collagen).
Enzymes act as BIOLOGICAL CATALYSTS, which means that they speed up a chemical reaction.
Without them, many processes in the body would occur too slowly. Before continuing, let us define three
important terms when it comes to chemical reactions in the body.
Metabolism All the chemical reactions that occur in the Respiration occurring in the
body. mitochondria of a cell.
The table below shows examples of a few enzymes you will learn in A’ Level Biology.
Amylase Breaks down starch into maltose. Secreted by salivary glands and the pancreas.
Pepsin / Trypsin Breaks down proteins into polypeptides and into amino acids.
DNA ligase Joins two pieces of DNA molecules together, such as during genetic
engineering.
As mentioned before, enzymes are globular proteins with 3D tertiary structures. They can either be
intracellular (inside the cell) or extracellular (outside of the cell, such as pepsin).
Enzymes bind with SUBSTRATE molecules to form an ENZYME-SUBSTRATE COMPLEX and finally
convert them into PRODUCTS. For a breakdown of starch, for example, starch would be the substrate,
amylase is the enzyme and maltase is the product. The enzyme is left unaltered at the end.
The substrates are always in motion due to their kinetic energy, so think of them as rapidly colliding with
the enzymes until they bind. The active site has R groups that interact with the substrate.
The substrate temporarily binds with the enzyme’s ACTIVE SITE, which is a specific shape on the
enzyme’s surface. This is often referred to as a LOCK AND KEY mechanism if it is a perfect fit.
However, some enzymes alter their shape slightly to accommodate holding the substrate in place.
This is known as INDUCED FIT. Think of how a glove may stretch slightly to accommodate a hand. The
diagram below shows this.
The graph shows the usual course of an enzyme reaction. With the enzyme, you will see that the initial
rate is very high. However, as a little time passes, it plateaus. Why is this? First, keep in mind there are
usually more substrate molecules than enzyme molecules.
At A, the substrates are rapidly binding with the available enzymes so the rate of conversion of substrate
to product is at its PEAK here.
At B, all of the enzymes are currently ‘occupied’ and as such, the rate of product formation DECREASES
as the substrates now must ‘wait’ for an enzyme active site to become free.
At C, there are very few substrate molecules left. Very little product remains to be formed now, so the
rate is very low (almost a plateau) until all of it has been converted.
The graph also shows that without the enzyme, the SAME AMOUNT of substrate would be converted
into product, but it would take a much longer time. This would also happen in a more LINEAR manner.
WHAT HAPPENS IF YOU INCREASE THE AMOUNT OF SUBSTRATE?
On the graph, you will notice that nothing has really changed in terms of how rate of reaction occurs
when the amount of substrate has been increased. It still begins rapidly, slows down and eventually
plateaus.
As before, more substrates with the same amount of enzymes means that the enzymes become quickly
‘occupied’. Other substrates would be rapidly colliding with the enzymes but would be unable to bind and
must ‘wait’ until one’s active site is free. The section marked Vm indicates that the enzyme is working at
its maximum possible rate, at full capacity.
Think of it as many people lining up to go into a building. They will eventually get in, but it will take a
while.
Recall that starch is broken down into maltose by the enzyme amylase.
In an experiment, imagine if the amount of starch substrate is in equal amounts in each trial. However,
what is being varied now is the amount of enzymes available. With the same amount of substrate but
more enzymes, the reaction will INCREASE. The INITIAL rate of reaction will increase
PROPORTIONATELY.
Think of it as people queueing up at a bank. However, more tellers have now opened up their stations and
now more lines can form. As a result, the transactions will occur at a much faster rate. In this analogy, the
‘people’ are STARCH. The ‘tellers’ are AMYLASE. The ‘transactions’ refer to the conversion of starch
to MALTOSE.
Recall that substrates have KINETIC energy in their molecules that allow them to rapidly move, collide
and eventually bind with enzymes.
If this energy is too low, they will move much more slowly and with much less momentum, so it is less
likely for them to bind. As such, the rate of reaction INCREASES as TEMPERATURE increases.
Reaction rate is actually said to DOUBLE every 10 C increase. This is called the Q TEMPERATURE
o
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COEFFICIENT.
SAMPLE GRAPH:
HOW DOES PH AFFECT ENZYME ACTIVITY?
Think of pH as the suppression of HYDROGEN IONS in a solution. So we can say that the lower the pH,
The graph shows the effect of pH on two proteases, pepsin and trypsin. Both perform the same function
(hydrolysing proteins into amino acids) but are found in different parts of the body. As a result, they both
have different optimum pH’s. Pepsin works best in an ACIDIC pH while trypsin works best in an
ALKALINE pH.
4.4: Explain the effects of competitive and non-competitive inhibitors on enzyme activity.
An INHIBITOR is a substance that will decrease the rate of an enzyme reaction, or stop it altogether. It
might do this by preventing the substrate from binding to the active site. Sometimes, inhibitors have very
similar shapes to the substrates and may bind to the enzyme instead of the substrate, ‘occupying’ the
space.
Many times, this is a REVERSIBLE process and does no damage to the enzyme or the active site and it
functions normally afterwards. Sometimes it can permanently alter the enzyme’s shape, thus preventing
any substrate molecule from attaching to it.
From the graph, it can be seen that with COMPETITIVE inhibition, INCREASING the amount of
substrate will raise the initial rate of reaction. This is because the enzyme is still functioning but the
substrate is temporarily blocked from the active site from time to time.
This is not so with NON-COMPETITIVE inhibition. Because the enzyme has been altered (reversibly or
irreversibly), increasing the substrate concentration does NOT increase the rate of reaction. It makes no
difference if the enzyme itself cannot function properly.
The table below shows some examples of competitive and non-competitive inhibitors:
Lead (heavy metal) Non-Competitive Breaks disulphide bonds in enzymes, denaturing them.
• Competitive inhibitors prevent substrate from binding to ACTIVE site. No significant change in
active site shape occurs but substrate is blocked. Non-competitive inhibitors bind to
ALLOSTERIC site and significantly changes active site shape while inhibitor is binded.
1.1: Illustrate the structure of RNA and DNA using simple labelled diagrams.
It is important to recall that DNA stands for DEOXYRIBONUCLEIC ACID and RNA stands for
RIBONUCLEIC ACID. This is because DNA lacks an OXYGEN that RNA has. They are mainly found
in the NUCLEUS of the cells and their tasks are to produce a genetic code to express certain traits, such
as eye colour, blood type and whether or not a disease is present, such as haemophilia.
The DNA has the shape of a DOUBLE HELIX. Each chain of this helix is made of NUCLEOTIDES,
which each have organic BASES that are connected by HYDROGEN bonds.
There are FOUR DNA bases, named ADENINE, CYTOSINE, THYMINE and GUANINE. Respectively,
these are represented as the letters A, C, T and G.
Base Type
Pairs With
A Purine T
G Purine C
C Pyrimidine G
T Pyrimidine A
1.2: Explain the importance of hydrogen bonds and base pairing in DNA replication
You may recall that when cell division occurs, this is called MITOSIS. Mitosis allows one parent cell to
divide into two identical (clone) daughter cells. When mitosis occurs, the DNA replicates. What this
means is that it produces TWO copies. Where does this other copy come from?
What exactly is happening here? Let’s make sense of this.
1. An enzyme known as DNA HELICASE ‘unzips’ the DNA into two strands by breaking the
HYDROGEN bonds between the bases. There is now a ‘leading’ and ‘lagging’ strand.
2. Another enzyme known as DNA POLYMERASE slides along the strands and pairs free
nucleotides with the ones attached to the original strands (for e.g. a free-floating C in the nucleus will
bind to the G on the original strand).
3. HYDROGEN bonds form, linking the two, and there are now two DNA molecules!
The concept was proven by two scientists named Meselson and Stahl. They submerged E. coli bacteria
in ammonium chloride with the nitrogen isotope being ‘dense’ (N-15). This meant that this isotope was
all the bacteria should’ve had N-15 in its DNA. The cells were then transferred to a medium containing
the isotope N-14, which is ‘less dense’.
The bacteria were harvested and the DNA collected and dissolved in caesium chloride (CsCl). This was
then put in a centrifuge and a concentration gradient was established. Observe the diagram below. After 1
generation, it showed a band of DNA of intermediate density.
DIFFERENCES BETWEEN DNA AND RNA
1.3: Explain the relationship between the sequence of nucleotides and the amino acid sequence in a
polypeptide.
A GENE is the length of DNA that codes for a single polypeptide. A tiny alteration in the sequence of the
DNA can cause a large change in the protein synthesized. If this occurs randomly (usually due to a
‘copying error’), it is called a MUTATION.
As shown in the image below, there are a number of processes that take place for the DNA to be coded
into an amino acid, which will comprise the protein. The two main ones are:
1. TRANSCRIPTION – The DNA code is copied onto a molecule called MESSENGER RNA
(mRNA). This is done three bases at a time, called a base TRIPLET or a CODON. Remember
that RNA does not have thymine (T) so adenine (A) on the coding strands is transcripted as
URACIL (U) on the mRNA.
2. TRANSLATION – The 3-letter codon serves as instructions for the formation of an amino acid
molecule by the RIBOSOME. Sometimes an amino acid may form from multiple codons, e.g. CCA, CCC
and CCG all form glycine. These are DEGENERATE codons.
WHAT IS PHENOTYPE?
Heterochromatin is denser, more tightly coiled and darker in colour. This is found in DNA not being used
for transcription.
The PHENOTYPE of an organism refers to the expression of a set of genes (e.g. black fur, blue eyes,
having a widow’s peak).
Recall that DNA determines the sequencing of the amino acids that produce the polypeptides and proteins
for these genes. Therefore, DNA highly influences phenotype.
It should be noted that phenotype can be influenced by the ENVIRONMENT. For example, genes can
code for expressions of light complexions. However, exposure to sunlight can stimulate MELANIN
production in the basal epidermis, giving that person a darker complexion.
2.1 & 2.2: Describe, with the aid of diagrams, the processes involved in mitotic cell division (including
interphase); AND observe freshly prepared root tip squash to show the stages of mitosis;
Role Explanation
Asexual When one parent cell splits into two identical daughter cells, this leads to asexual
reproduction reproduction in many organisms, both unicellular (such as AMOEBA) and
multicellular (such as in BRYOPHYLLUM meristems or in HYDRA). This is
because there is only ONE parent.
Growth Since mitosis results in cloned daughter cells, growth can normally occur in areas
such as the MERISTEMS of plants or of the ZYGOTE of animals, which will keep
dividing to eventually form the embryo.
Tissue repair Mitosis is used to regenerate any cells or tissue that has been lost due to damage or
age. The cells will divide and form new cloned daughter cells.
Immunity When exposed to pathogens, the body will ensure that there are adequate
LYMPHOCYTES to produce sufficient antibodies to destroy the foreign invaders.
When one is ill, there is always an excess production of white blood cells.
2.5: Explain what is meant by homologous pairs of chromosomes, and the terms haploid and diploid.
2.6 & 7: Describe with the aid of diagrams, the processes involved in meiotic cell division, and describe
how meiosis contributes to heritable variation.
Meiosis is type of cell division, in ways similar to mitosis, but with many key differences, especially
concerning the daughter cells that are formed from the parent cell. Meiosis occurs in any organisms that
undergo SEXUAL reproduction and must produce sex cells or GAMETES.
When two gametes fuse during FERTILIZATION, they form a ZYGOTE and be the first cell of a new
organism. This cell then repeatedly divides by MITOSIS as the organism undergoes growth and further
development.
Meiosis is mainly different due to the fact that it produces daughter cells with GENETIC VARIATION,
meaning each is different from the parent and from each other. This is due to DNA CROSSING OVER.
Chromosomes arrange themselves in pairs called BIVALENTS and cross at CHIASMATA. Meiotic
daughter cells also only have HAPLOID numbers, meaning they have half the number of chromosomes.
There are THREE main mechanisms involved in sexual reproduction that contribute to genetic variation.
Two of these occur during meiosis and the third occurs during fertilization.
Mechanism Explanation
INDEPENDENT Chromosomes pair up into bivalents and align at the equator of the cell. One
ASSORTMENT from this pair come from the mother and the other from the father. These pairs
are each sorted into the daughter cells independently and can result in a
massive number of combinations.
CROSSING OVER Previously mentioned, crossing over in the bivalents occur in fusion and
breakage points in the bivalent chromatids called chiasmata.
RANDOM Remember that gametes have a haploid number of chromosomes, which are
FERTILIZATION all independently assorted and crossed over. Each gamete is genetically
unique. During sexual reproduction, the gametes that fuse are up to chance.
Reason Explanation
Limiting spread of disease Genetic variation ensures that each member of a species has varying
levels of immunity against communicable diseases. If all the organisms
were genetically uniform, diseases would spread very quickly through
populations.
A popular example involves the fungal Panama disease against Gros
Michel banana cultivars.
Ensuring a species can Genetic variation ensures that individuals are diverse enough to be able to
adapt to environmental survive in different climates, temperatures and differences in abiotic
changes factors. With climate change on the rise, vegetatively propagated plants
that cannot adapt to high fluctuations in temperature may die.
Preventing If organisms are diverse, so would their needs for survival such as diet
overcompetition between and habitat. Due to this, these species would not have to fight for limited
members of the species. resources.
The basic example of this are the finches on the Galapagos Islands,
studied by Charles Darwin to form his theory of evolution.
It is important to note that meiosis occurs in TWO main stages that are simply named MEIOSIS I and
MEIOSIS II. Meiosis I involves independent assortment and crossing over, while the stages in Meiosis II
is almost identical to mitosis.
If a cell dies, the body must replace that cell. The only way to replace the cells is to first copy the
information that the cell contained. There is a complex system of proteins and enzymes that unravel the
DNA double helix so that the DNA can be copied.
If a single cell dies it can be replaced through MITOSIS. This system works well with single cell and
simple organisms. More complex organisms use meiosis to produce gametes (egg or sperm cells) for
sexual reproduction. Meiosis also begins with DNA replication. The genetic stability of a multicellular
organism is reliant on an accurate DNA replication system.
Sometimes, a MUTATION may occur. This occurs when there is a RANDOM and unpredictable error in
this copying system. This can occur in numerous ways, such as a base being deleted, substituted or an
extra base being added. Sometimes MUTAGENS, such as carcinogens of high-frequency radiation, can
increase the likelihood of mutations by damaging the DNA.
Note that mutations can be categorised into two main groups: GENE and CHROMOSOME. There are
also numerous ways in which gene mutations can occur:
SUBSTITUTION (or Replaces one base with another. Sometimes this does not
POINT MUTATION) result in any change (as many triplets code for the same
protein), so it is often called a SILENT mutation.
Examples include SICKLE CELL ANAEMIA and PKU.
DELETION The loss of a base pair. Changes how the entire DNA
sequence is read. Also called a FRAME SHIFT
mutation, due all the bases being ‘shifted’.
While gene mutations are changes in the nucleotide sequences in the DNA (such as by insertion,
substitution or deletion), chromosome mutations are changes in the cell’s chromosome NUMBER or
chromosome STRUCTURE.
When the cell divides during the first phases of MEIOSIS, there is a random chance of the chromosomes
being pulled apart unevenly between the daughter cells. When this happens, it is called NON-
DISJUNCTION.
HOW IS MUTATION RELATED TO GENETIC VARIATION?
Recall that all members of the same species are able to interbreed and produce FERTILE offspring. Every
species has some genetic variation, due to only half of the chromosomes being passed down from each
parent after being independently assorted and crossed over. Note that ENVIRONMENTAL variation
(such as sunlight exposure affecting skin complexion) is not passed down through genes.
A MUTATION is a random change in the DNA, sometimes involving a trait not present in the parent
organism. As a result of this, new PROTEINS may be translated from the new nucleotide sequences,
resulting in new PHENOTYPES. This can have a great impact on NATURAL SELECTION, if these new
phenotypes give the organism an advantage over others in its environment.
3.1 & 2: Explain the terms: gene, allele, dominant, recessive, codominant, homozygous and heterozygous.
Use genetic diagrams to solve problems involving monohybrid and dihybrid crosses.
Term Definition
Allele A different form of a gene, found on the same locus of the chromosome.
‘Wild type’ alleles refer to those found naturally in populations.
Dominant Describes an allele that will express its trait even if a different allele is present.
Recessive Describes an allele that will only express its trait if a dominant allele is absent.
Codominance Describes alleles that produce a combined effect when expressed together.
Genotype A gene combination that will express a trait. (e.g. FF, Ff and ff are all genotypes)
Phenotype The observable characteristics expressed by that trait. (e.g. round or wrinkled
seeds).
Ultimately determined by genes, which are sequences of DNA that lead to the
formation of proteins. Changing a gene can thus change expression and phenotype.
Homozygous A genotype where both alleles are the same. (e.g. FF or ff)
Autosomes Refers to chromosomes that are not sex chromosomes (the first 22 pairs).
Dihybrid The inheritance of two genes at the same time (e.g. AABB, AaBb, aaBB, AAbb,
inheritance etc.)
Epistasis The event where the genotype for one gene affects the expression of another gene.
Chi-square (χ )
2
A statistical test that determines whether or not observed ratios are significally
test different from expected ratios.
Null hypothesis A statement in a chi-square test that says that there is no significant difference
between what is observed and expected.
When a single gene is inherited at a time, this is called monohybrid inheritance. Most of the Punnett
squares you’ve done previously has been related to this type of inheritance. Many traits, such as blood
type and eye colour, and inheritance of mutant alleles that cause diseases come as a result of this.
Let’s use CYSTIC FIBROSIS (CF) as an example. This is an inheritable disease that causes the body to
produce large amounts of thick mucus in the lungs and pancreas.
This mucus becomes a breeding ground for bacteria, which leads to other infections. CF is caused by a
‘faulty’ allele, which would usually produce a channel protein called CFTR, which allows flow of
CHLORIDE ions in and out of a cell.
The faulty allele ensures that the protein is not produced and chloride ions build up, resulting in the
mucus build-up. CF is inherited in an AUTOSOMAL RECESSIVE manner, meaning that the disease
only occurs if both RECESSIVE alleles are present in the genotype (ff). The presence of a dominant allele
(F) prevents the expression of the faulty recessive allele.
• Let’s observe how two parents who are carriers for CF can produce a child with CF.
WHAT ARE SEX-LINKED TRAITS?
Sex-linked (not to be confused with sexually transmitted) traits occur when the alleles are placed in either
the X or Y chromosomes. Note that these two chromosomes are not HOMOLOGOUS, which means that
they would not have the same number of gene loci.
The Y chromosome is small compared to the X and has less available positions for alleles, so some alleles
will be present in the X but not the Y.
It is notable that men cannot pass on an allele from their sole X-chromosome to their sons as their sons
would always inherit their Y chromosome and the X from the mother.
One popular example of a sex-linked disease is HAEMOPHILIA, which occurs due to inheritance of a
faulty allele that is placed on the X chromosome but which locus is absent on the Y chromosome.
Therefore, if a boy only has one faulty allele, he will have haemophilia. A girl would need two faulty
alleles. As a result, it is more likely for boys to inherit haemophilia.
Using X as the normal blood clotting allele and X as the haemophilia allele, complete the Punnett square
H h
below. (Remember the Y chromosome does not have a locus for the allele, so it’s left blank.)
Genotype Phenotype
RRYY Round, yellow peas
RRYy Round, yellow peas
RRyy Round, green peas
RrYY Round, yellow peas
RrYy Round, yellow peas
Rryy Round, green peas
rrYY Wrinkled, yellow peas
rrYy Wrinkled, yellow peas
rryy Wrinkled, green peas
As previously said, sometimes during dihybrid inheritance, the expression of a pair of alleles can have
influence over the expression of others. Sometimes a trait will not manifest because of the expression of
another trait, or if a pair codes for the absence of a certain enzyme. A very common example occurs with
coat colour in animals. Coat colour depends on the presence of pigments.
However, if a pair of alleles determines the organism does not have a pigment (an ALBINO), then the
other allele combinations would not matter. If the first pair of alleles determines the animal is an albino,
then no other coat colour is even possible.
This is known as EPISTASIS. It should be noted that epistasis can be either dominant or recessive.
Let’s look at this example below. Picture a species of mouse that can have either a brown (b) or black (B)
alleles for coat colour. Black is dominant to brown. However, another pair of alleles code for the presence
of melanin to produce the coat. ‘C’ represents melanin production and ‘c’ represents no melanin.
WHAT IS THE χ (CHI-SQUARE) TEST?
2
The χ2 test is a statistical test that is often used to compare OBSERVED results with EXPECTED results
to determine if there are any significant differences between them. This is done to see if there are any
external variables affecting the results, such as environmental factors, mutations or human intervention.
To do any statistical test, a NULL HYPOTHESIS is first set up. A null hypothesis would read as:
H : The observed results are not significantly different from the expected results.
0
If this is not so, then the ALTERNATIVE HYPOTHESIS must be accepted, which would read as:
H : The observed results and expected results have a significant difference between them.
1
QUESTION ONE: “During dihybrid inheritance, two pea plants of genotypes RrYy were crossed. This
should have yielded a 9:3:3:1 ratio. The offspring were counted and the numbers were recorded.”
The degrees of freedom would be 3. Use the table on the previous page to determine whether to accept the
null hypothesis or alternative hypothesis.
Sum (χ value) =
2
13.36
In this case, the ALTERNATIVE hypothesis would be accepted, meaning that there IS a significant
difference between the observed and expected results. We have rejected the null hypothesis.
QUESTION TWO: “During dihybrid inheritance, brown coat and black coat mice were mated and their
offspring observed. They experienced recessive epistasis, meaning some offspring had white coats. Their
expected ratio was 3 brown : 3 black : 2 white.”
Use the table on the previous page to determine the ‘p’ value.
Phenotype Observed (O) Expected (E) (O – E) (O – E) 2
(O – E) / E
2
Sum (χ value) =
2
0.77
In this case, the NULL hypothesis would be accepted, meaning that there ISN’T a significant difference
between the observed and expected results.
4.1 & 4.2: Outline the principles of restriction enzyme use in removing sections of the genome, and
explain the steps involved in recombinant DNA technology.
Genetic engineering (GE) can be defined as THE ALTERING OF THE DNA in an organism, usually by
extracting and inserting the DNA from a member of one species to another species. This altered DNA is
called RECOMBINANT DNA and the organism which genome now contains it is called a GMO, which
is short for GENETICALLY MODIFIED ORGANISM.
It is often compared to ARTIFICIAL SELECTION (or selective breeding), but they remain two entirely
separate processes. Whereas both involve the passing of traits from one organism to another, GE is a
more specific and expensive process, using enzymes to cut, transfer and attach DNA, one gene at a time.
It can also change the DNA of the organism without having it inherit the traits from its parents. This
means it can be used to treat certain diseases, such as SICKLE CELL ANAEMIA.
So what happens?
• First, lengths of DNA are cut from the human DNA. This is done using a RESTRICTION
enzyme. These are made by bacteria to fight off bacteriophage viruses.
• Similarly, the restriction enzyme makes cuts on a bacteria PLASMID (a circular length of DNA).
Imagine that this leaves a gap in that DNA. This is also called a VECTOR.
• Another enzyme named DNA LIGASE is used to join both sets of DNA to make a segment of
RECOMBINANT DNA.
• The recombinant DNA is inserted into the bacteria, which now becomes a GMO. The
recombinant bacterial cells are now cloned. This can be done using a process called PCR.
• This process is just a general overview. There are many variations and processes involved.
NOW LET’S GET MORE SPECIFIC... How is the gene isolated?
INSULIN is a hormone (a protein) that is produced by the BETA cells of the human PANCREAS. It is
necessary for the conversion of GLUCOSE TO GLYCOGEN. GE is used in the modern day to produce
large amounts of insulin from transgenic E. coli. The first step is to extract the human DNA code for
insulin production.
This is done using an enzyme called REVERSE TRANSCRIPTASE. This enzyme takes the mRNA
extracted from the pancreatic beta cells and uses it as a template to produce a single complementary
strand of DNA.
DNA POLYMERASE is now used to attach free nucleotides to form the other strand. The insulin gene
has now been isolated.
A PLASMID is commonly used as a vector to transport genetic material from one organism to another.
Recall that plasmids refer to DNA packaged in a circular form and are usually found in bacterial cells.
Another thing to note is that plasmids often contain some genes that make them resistant to
ANTIBIOTICS.
When bacteria are attacked by viruses, they produce RESTRICTION ENZYMES. These can be used to
cut particular base sequences of DNA. A popular example of a restriction enzyme is EcoRI, which cuts a
GAATTC sequence (as well as cDNA sequence).
Observe that the cuts are asymmetrical and so leave points that jut out. These are known as STICKY
ENDS and can easily form hydrogen bonds with complementary base pairs and form new DNA. This is
also done to the insulin DNA. The bacterial plasmids and the insulin DNA are then mixed until they pair
with each other to form RECOMBINANT DNA. An enzyme called DNA LIGASE is then used to ‘tie’
everything together, linking the sugar-phosphate backbones of the DNA molecule.
The recombinant plasmids are mixed in a solution containing the bacterial culture. However, most of the
bacteria do not get the plasmids into their cells. Only about 1% actually take the plasmid into them.
So how do we know which ones are of that 1%? It so happens that when the plasmid was altered before
fitting it with the insulin gene, another gene was inactivated. This gene provided ANTIBIOTIC
RESISTANCE for an antibiotic named TETRACYCLINE.
Let’s recap some of the important factors needed:
Compound Role
RESTRICTION Used as “molecular scissors” to cut pieces of DNA. Leaves sticky ends to
ENZYMES attach complementary pieces of DNA.
REVERSE Synthesizes DNA molecules from mRNA template.
TRANSCRIPTASE
DNA LIGASE Used as “molecular glue” to connect lengths of DNA and form bonds.
DNA POLYMERASE Used in DNA replication, to produce a complementary strand of DNA
from a single DNA molecule.
4.3: Discuss the successes and challenges of gene therapy in modern medicine.
Gene therapy is the process of TREATING OR PREVENTING DISEASE BY ALTERING THE GENES
IN A PERSON’S CELLS.
There are numerous success stories and challenges associated with gene therapy:
SCID: The first human to receive gene therapy was an infant girl with SCID (severe combined immune-
deficiency), which is caused by a faulty gene that prevented the production of an enzyme. Due to this, the
girl was unable to fight off PATHOGENS. The correct allele was inserted into the girl’s white blood cells
using a RETROVIRUS vector and re-inserted into her body. She was then able to resist pathogens and
lead a normal life.
CYSTIC FIBROSIS (CF): Patients with CF tend to produce thick MUCUS in multiple parts of the body.
This mucus can become breeding grounds for bacteria and thus, many organs may be prone to infections.
It may even cause men to be sterile due to blocking ducts in the male reproductive system.
CF is caused by a defective recessive allele that is responsible for a carrier protein called CFTR. The
protein is not placed on the plasma membrane. Researchers first placed the correct allele in a
LIPOSOME, which could diffuse through the phospholipid bilayer. It worked only temporarily.
Researchers then tried VIRUS vectors to transport the gene but patients began to suffer side-effects from
infection. The studies are ongoing.
With the advent of technologies such as CRISPR-Cas9, studies are currently being done on treating
diseases such as: SICKLE CELL ANAEMIA, HAEMOPHILIA, AIDS and LYMPHOMA.
4.4: Discuss the implications of the use of GMO’s on humans and the environment.
Type of Details
implication
Environmental • The release of a GMO species would have the possibility of causing an
ECOLOGICAL IMBALANCE. The main concern is that crops can spread
their genes to wild plants through pollination.
Medical • It is possible for ALLERGENS can be transferred from one food crop to
another through genetic engineering. Another concern is that pregnant
women eating GMO products may endanger their offspring by harming
normal fetal development and altering gene expression.
• GMO’s may lead to diseases not yet known. As defective genes are replaced
with functional genes, it is expected that there will be a reduction in genetic
DIVERSITY, making the population more susceptible to infections.
5.1 – 5.3: Explain how environmental factors act as forces of natural selection; how natural selection
may be an agent of change or constancy; and how it is a mechanism of evolution.
Only the organisms BEST ADAPTED are the ones more likely to survive and pass on these advantageous
traits to their offspring. The species eventually becomes better and better adapted to overcome these
selective pressures.
E.g. if members of a species develop immunity to a certain infectious, deadly disease, these are the ones
that will survive and reproduce.
Observation Deduction
Members of a species VARY between If traits can be inherited then the organisms will pass them on
each other. Some of these variations to their offspring.
are INHERITED.
Population numbers remain fairly Members that are BEST ADAPTED to their environment are
CONSTANT over long periods of the ones most likely to survive, reproduce and pass on their
time. ADVANTAGEOUS traits.
Observation Deduction
TRINIDADIAN GUPPIES Trinidadian guppies are models of natural selection. They have
developed various mechanisms that help them evade predators.
They can produce excess PIGMENT in their eyes, making them
black, to throw off predators’ aim.
Depending on their predators, they will grow to different SIZES
upon sexual maturity. For e.g. guppies with cichlid predators
grow to smaller sizes since cichlids tend to hunt larger fish.
CARIBBEAN ANOLE Anole lizards have been observed to branch into many different
LIZARDS colours and sizes, called ECOMORPHS. This is totally
dependent on their habitat and diets, just like with Darwin’s
finches.
It was observed that a single type of anole lizard in each island
branched into the same ecomorphs on the different islands,
providing evidence that evolution can be predicted and repeated.
This, along with the guppies, have also shown evolution
occurring on a much shorter timespan.
THE CASE OF THE PEPPERED MOTH (Biston Betularia)
Antibiotics are chemicals that are ingested to kill bacteria. They are usually produced by other living
organisms. A prime example is PENICILLIN, which is produced by the Penicillium fungus. Penicillin
prevents the formation of CELL WALLS in bacteria cells. However, some MUTANT bacteria have
produced an enzyme which inactivates penicillin and thus, have become RESISTANT to it.
As a result, any bacteria that isn’t resistant to penicillin will die, leaving the mutant population to
REPRODUCE and rapidly increase. This is highly dangerous, especially to patients who are already very
ill. The antibiotics in this case would be an example of a SELECTIVE PRESSURE. There exists a type of
bacteria called MRSA (methicillin-resistant Staphylococcus aureus) which are resistant to numerous
antibiotics and usually infect patients with compromised immune systems.
There is also the case of INSECTICIDE resistance, where many pests have grown resistant to them. This
is usually an argument for BIOLOGICAL CONTROLS being used instead of insectides.
WHAT ARE THE DIFFERENT TYPES OF SELECTION?
Observation Deduction
At O’ Level, you would have learnt that two members of the same species:
This is known as the BIOLOGICAL SPECIES CONCEPT and has been used to classify organisms into
different taxa quite successfully, with each species being given their own binomial name, e.g. Canis
lupus, Ursus arctos, Leo panthera and Homo sapiens.
There exists one major limitation with rigidly sticking with this concept, however: It only applies to
organisms that reproduce SEXUALLY. It also cannot be used to classify organisms that are EXTINCT.
This is where an alternate method exists called the PHYLOGENETIC SPECIES CONCEPT. In this
method, organisms are classified according to certain defining traits or MORPHOLOGY. This is used to
trace a ‘genetic history’ of the organism, tracing back to its common ancestors.
One major limitation with this method, however, is dealing with species that demonstrate
POLYMORPHISM, or having many different forms that can be mistaken for different species. Both
white and dark variants of the peppered moth, for example, could easily be mistaken as two separate
species just by looking at them.
Extinct species Cannot be used to classify extinct Can be used to classify extinct species and
species and fossils. fossils.
WHAT IS SPECIATION?
Speciation, put simply, is the formation of a new species or when two or more species branch out from a
common ancestor. This usually occurs due to a mechanism known as ISOLATION, which sets up
different kinds of barriers that prevent members of a species from interacting. These barriers can be
geographical, behavioural or even based on times of sexual maturity.
When isolation occurs, various groups of the same species may be subjected to different SELECTIVE
PRESSURES and would have to ADAPT to varying circumstances. After a while, no GENE FLOW will
occur between the splintered populations and each population will evolve into a new species.
GEOGRAPHICAL Occurs when two species are Darwin’s finches were separated from each
PHYSICALLY SEPARATED by other by living on different islands in the
a large mass, such as an ocean or Galapagos. Gene flow was not possible.
mountain. Leads to ALLOPATRIC speciation.
ECOLOGICAL Occurs when two species live in Red-legged frogs and American bullfrogs
the same area but RARELY OR live in the same ecosystem. However, red-
NEVER MEET. legged frogs dwell in streams while
bullfrogs breed in ponds.
BEHAVIOURAL Occurs when two species have Eastern and western meadowlark birds have
different COURTSHIP different mating calls. Different species of
behaviours. fireflies have different lighting signals.
MECHANICAL Occurs when the two species are It is unlikely for white sage and black sage
incompatible, either in terms of to form hybrids in the wild as they are
size, genitalia or GAMETES. pollinated by two different types of bees.
TEMPORAL Occurs when two species live in Some cicadas tend to have 13-year cycles
the same place but experience while others have 17-year cycles. It is rare
different TIMES of sexual that these sync up for them to mate.
maturity.
ALLOPATRIC SPECIATION
Recall GEOGRAPHICAL isolation, where species will form splinter groups due to a separation by a land
mass, such as a mountain. The presence of this mountain means that the groups will not meet and thus,
will not mate. On either side of the mountain, there may be different selective pressures, such as different
coloured trees, terrain, predators, rainfall. Each member must now adapt to these pressures. This
geographical barrier will thus cause new species to arise.
• DARWIN’S FINCHES being physically separated by the water masses between islands.
• The PORKFISH and PANAMIC PORKFISH being separated by the isthmus of Panama.
• The KAIBAB, a subspecies of the ABERT squirrel live on opposite ends of the Grand Canyon.
SYMPATRIC SPECIATION
Whereas allopatric speciation deals with speciation arising in two geographically separated areas,
SYMPATRIC SPECIATION occurs when there is no geographical separation. If organisms living in
the same habitat, for example, can somehow overcome their temporal, ecological or behavioural barriers,
they may undergo sympatric speciation.
Examples include:
1.1 & 2: Describe the structure of the anther and the formation of pollen grains; and the structure of the
ovule and formation of the embryo sac
Flowering plants are also called ANGIOSPERMS. They contain reproductive organs, which produce sex
cells called GAMETES, similar to animals. These gametes, both male and female, are HAPLOID in
nature, meaning they contain half the number of chromosomes.
Male gametes are formed within the POLLEN grains, found in the ANTHERS, while female gametes are
found within EMBRYO SACS, found inside of the OVULE.
Reproductive Parts Comprises of Notes
As previously stated, the male gametes are formed inside pollen grains. These pollen grains are formed
from MICROSPORANGIAL cells found within four pollen SACS in the ANTHER of the flower.
Observe the micrograph and table below for the placement and roles of the various structures within the
anther:
Fibrous Layer Thickened cellulose walls. Eventually separates to release pollen grains.
Tapetum Inner layer of pollen sac that provides nutrition to developing grains.
Stomium The point at which dehiscence occurs, to release the pollen grains.
Pollen mother cell Diploid. Divide by meiosis to produce four haploid gamete nuclei.
Formation of pollen grains occur when a POLLEN MOTHER CELL undergoes MEIOSIS to form a
TETRAD of haploid cells called MICROSPORES. These then undergo MITOSIS to form two types of
nuclei within each, eventually maturing to form the protective walls of the pollen grain.
Antipodal cells Nourishes the embryo sac and endosperm. Located at chalaza, opposite to synergids.
Recall that in the pollen grains in the anther, there were TWO nuclei. One was the GENERATIVE
nucleus and the other was the TUBE nucleus. The generative nucleus divides by mitosis into two haploid
male GAMETES, while the TUBE nucleus allows the growth of a structure called a POLLEN
TUBE.
That might have been a lot to take in! So let’s recap the various parts involved:
Primary nucleus 2n Formed in embryo sac after two haploid nuclei fuse. In
centre.
Egg cell n Found among synergids in embryo sac. The female
gamete.
1.4 & 1.5: Explain how cross-fertilization is promoted; and genetic consequences of sexual reproduction
There are numerous reasons why some flowers or florists will want to allow self-pollination and self-
fertilization to occur (also called INBREEDING), as desirable traits can be predictably passed down
along generations and the flowers can be produced in very large numbers at a rapid rate. This happens
very easily with HERMAPHRODITIC plants, which mean they have both male and female parts.
Dioecious plants Male and female flowers of the same species are found on Chenet, paw-paw,
separate plants, making self-pollination difficult. marijuana
Monoecious plants Male and female flowers are located on the same plants but Pumpkin, maize,
may not mature at the same time, or be positioned to self- castor oil
pollinate.
Protandry and Either stamens mature before the stigmas (protandry) or Protandry –
protogyny stigmas are receptive to pollen before release (protogyny) Fireweed
Protogyny –
Soursop, avocado
Self- The pollen with the same genetic code as the stigma of the Tobacco, cabbage
incompatibility same flower will not germinate if contact is made.
Heterostyly The various forms of the flowers makes it difficult for self- Primrose, red
pollination to occur due to stigma and anther position. See cordia
previous page.
1.7: Discuss the development of the seed and the fruit from the embryo sac and its contents.
1. The zygote begins dividing by mitosis. A smaller TERMINAL cell is formed, with the initial zygotic
cell being called the BASAL cell. Surrounding the zygote is the ENDOSPERM, which nourishes it
and allows development to occur.
Egg cell Becomes a ZYGOTE (2n) and then undergoes mitosis to become an EMBRYO.
When this occurs, multiple structures develop such as a root (RADICLE), embryo
shoot (PLUMULE) and first leaves (COTYLEDONS).
Integuments Becomes the seed coat or TESTA as it becomes thickened and waterproof with lignin
and cellulose.
Petals Wither and fall off, along with the sepals. Exceptions include dandelions.
Endosperm Becomes the ENDOSPERM after mitosis, which nourishes the embryo during
nucleus germination, usually high in proteins and starches.
Ovary and The ovary becomes the FRUIT and the wall becomes the PERICARP, the ‘flesh’ of
ovary wall the fruit. The pericarp usually has some role to play in seed dispersal.
After fertilization occurs, the embryo sac and the ovule begin to undergo numerous changes as the ovary
becomes a fruit and the ovules become seeds. The table below will list some of those changes:
Asexual reproduction is defined as the production of new offspring by ONE parent through the process of
MITOSIS. The offspring are genetically identical to their parents or CLONES.
It is important to distinguish asexual reproduction from self-pollination, as the latter involves the
production and fusion of gametes from male and female parts and is thus considered SEXUAL
reproduction.
Advantages Disadvantages
Offspring remain well-adapted to a non- Due to lack of genetic variation, pathogens and
changing environment as all traits are inherited diseases may be able to spread very quickly through
from parent organism. populations.
Rapid growth of population can occur since Overcompetition may occur, either among offspring
only one parent is needed and no gestation or between parent and offspring, especially in plants
period. New habitats can be colonized quickly. due to being in close proximity.
Offspring may be able to utilize parent as a Very low genetic diversity can lead to lack of
nutrient source during the early stages of life. evolutionary changes in species.
Vegetative propagation is the process of producing plant offspring on a large scale, taking advantage of
the facets of asexual reproduction. This is usually done for commercial purposes and can be done with the
use of cuttings and tissue culture.
Cuttings
Cuttings are usually done for crops such as sugar cane. Stems are broken off and lain horizontally on soil.
After a period of time, buds grow into new stems and adventitious roots grow from the leaf scars. It
happens quickly as no POLLINATING AGENT is needed, so the crop is quite profitable.
For other plants (African violets, for example), the cutting is made on the stem and is plced in a medium
containing a growth hormone such as AUXIN, which stimulates root growth. The plant can then be
transferred to soil.
Tissue culture
Tissue culture is more frequently used in large scale production and can be done in a laboratory at any
location. Plant tissue is differentiated from animal tissue in that they are able to produce other cell types
(similar to stem cells). Because plant tissue can do this, they are said to exhibit TOTIPOTENCY.
A meristematic clump of cells called an EXPLANT is removed from the parent and placed in a STERILE
nutrient solution (usually high in sucrose, Vitamin B, nitrates and mineral ions) containing AUXIN and
CYTOKININ, hormones that stimulate cell growth and division. Sterility is key to prevent infection by
pathogens and fungi. The explants then undergo mitosis to form larger clumps called CALLUSES. When
the plant is at a certain point of development, they are transplanted into sterile soil.
2.1&2: Describe the structure & function of the male & female reproductive systems; and gametogenesis
Stage Description
Ovulation The release of a secondary oocyte (not ovum) from the ovary.
Copulation / Coitus The act of intercourse, where male gametes are delivered to the female gametes.
Fertilization The fusion of the nuclei of the male and female gametes.
Implantation The action of the early embryo sinking into the endometrium.
Gestation The period between conception and birth, where foetal development occurs.
Recall that for gametes to be produced, diploid cells must divide by meiosis to produce haploid cells. So
Spermatogenesis begins at a layer of cells called SPERMATOGONIA placed along the outer wall (called
the germinal epithelium). These divide by mitosis and grow into larger structures called PRIMARY
SPERMATOCYTES.
The diploid primary spermatocytes divide by MEIOSIS to now to first become haploid SECONDARY
SPERMATOCYTES and then into SPERMATIDS. Spermatids will then grow a tail-like structure as they
specialize to become SPERMATOZOA, the male gametes.
There are a few idiosyncrasies when it comes to the production of female gametes (oogenesis). You may
have learnt at O’ Level that the egg cells (or ova) are released from the ovaries during ovulation. This is
not entirely true as the actual structure that is released is a precursor to the ovum, called a SECONDARY
OOCYTE.
OOGENESIS
In order to understand what is happening during oogenesis, we should look at a cross-section of the ovary.
There are notable similarities and differences when compared to spermatogenesis. For example, haploid
cells are produced during meiosis and the process begins along germinal epithelial cells in the ovaries.
However, this begins when the girl is an embryo instead of at puberty, like a boy.
2.3: Discuss how the structure of the ovum and the sperm facilitate their functional roles in fertilization.
First, let’s recap some facts and comparisons about spermatogenesis and oogenesis.
Timeline Starts at puberty and continues into Starts when female is a foetus, then stops.
old age. Uninterrupted until death. Resumes at puberty and an egg is released
monthly. Terminates at menopause.
Production About 200 million sperm daily, Releases one secondary oocyte every menstrual
rate fully matured. cycle.
GAMETE STRUCTURE
Aspect Spermatozoa Secondary oocyte
Size Head is about 3µm wide and tail is About 100µm in diameter. Much larger.
50µm long. Much smaller.
Membranes Plasma membrane around head with Has multiple layers: a plasma membrane with
glycoproteins that support union with microvilli, glycoprotein-rich zona pellucida and
oocyte. corona radiata.
Meiotic Already completed meiosis II upon Only completes meiosis II after fertilization to
stage release. become ovum.
2.4: Discuss the basic process of fertilization.
FERTILIZATION
The basic definition of fertilization is the fusion of the nuclei of both male and female gametes.
Fertilization takes place in the OVIDUCT. Sperm are ejaculated during coitus, caused by the stimulation
of nerves along the vasa deferentia and muscular contractions push sperm out of the urethra. The sperm
use semen as a medium, a fluid containing CALCIUM ions, CITRATE and FRUCTOSE.
2.5, 2.8 & 2.9: Discuss the process of implantation; the structure & functions of the placenta and amnion.
IMPLANTATION
When the zygote forms, it continuously divides by MITOSIS to form a ball of cells known as a
BLASTOCYST. The blastocyst moves along the Fallopian tube due to contractions along its muscular
wall and with help from cilia. It will finally reach the uterus, where it will attach itself to the lining of the
uterus wall, or ENDOMETRIUM. It is now said to have been implanted.
Nutrient and CHORION facilitates diffusion of glucose and amino acids. Active transport of
antibody intake ions. Nutrients stored in YOLK SAC.
2.6: Discuss the importance of hormones in gametogenesis and the menstrual cycle.
Before we discuss the intricate systems that these hormones belong to, let’s summarize them:
Inhibin Testes Inhibits the release of GnRH, thus also inhibiting release of
FSH and LH.
DMPA and Synthetic Prevents ovulation or thickens cervical mucus. Used for birth
Progestin control.
HORMONAL CONTROL OF GAMETOGENESIS
Spermatogenesis
Oogenesis
• Day 6 – 14 – FOLLICULAR PHASE – The presence of FSH and LH triggers the release of
OESTROGEN from the ovarian THECA. As oestrogen levels rise, this causes a positive feedback
effect and a surge in LH. Oestrogen levels plummet as GnRH is inhibited.
• Day 14 – This surge in LH triggers OVULATION, causing the follicle to rupture and secondary
oocyte to release. The ruptured follicle becomes a CORPUS LUTEUM.
• Day 14 – 28 – LUTEAL PHASE – The corpus luteum secretes PROGESTERONE to keep the
uterus lining thick for implantation. If pregnancy does not occur, the corpus luteum decays into a
scar called a CORPUS ALBICANS. Oestrogen and progesterone levels decrease.
• Day 0 – 6 – MENSES then occurs as the uterus lining sheds. The drop in the gonadal hormones
causes a slight increase in FSH and LH, restarting the cycle.
2.7: Discuss how knowledge of human reproductive anatomy and physiology has been applied to the
development of contraceptive methods;.
Birth control, you would have learnt, involves methods of preventing pregnancy from occurring. We can
class birth control into two categories:
Barriers (e.g. They create an impermeable physical Most popular method, though some say
condoms, femidoms barrier that prevents sperm cells from they reduce pleasure. Also prevents
and diaphragms) entering the vagina or uterus. As a transmission of STI’s. More effective
result, no contact can be made with when used with a SPERMICIDAL
the egg. CREAM.
Progestin implants A rod-shaped device is implanted in Does not protect against STI’s. Works
the uterus. It releases synthetic similarly to birth control pills.
progestin into the blood, which
inhibits ovulation.
Oral contraceptives Usually contain synthentic oestrogen The “mini-pill” thickens cervical mucus,
(birth control pills) and progesterone to mimic preventing the sperm from entering.
pregnancy and suppressing
ovulation.
Sterilization The vasa deferentia of the men are Is 100% effective, so patients must be
cut and tied, preventing sperm from certain they want no more children.
entering the urethra. Or the oviducts
are cut and tied (tubal ligation).
Filshie clips A device that is clipped across each Reversal of ligation more likely to be
oviduct during tubal ligation. successful when this is implemented.
But there are infectious risks if the clip
opens or migrates.
Emergency Various types exist. The mechanism Despite its name, many of them can be
contraception or the of action usually involves delaying taken from 3 – 5 days after and certainly
“morning after” pill just in the morning. However, success
ovulation or causing changes in the increases when it is taken closer to
endometrium that limit implantation. period of coitus.
Intra-uterine A T-shaped device made of copper Copper is toxic to the sperm as well, so
devices (or IUD’s) and plastic that is inserted into the it can be seen as a contraceptive method
uterus by a physician. It stimulates as well.
immune responses in the uterus to
attack the sperm or embryo.
There are usually ethical debates about birth control. The table below summarizes some of the arguments
that are pro and con:
For Against
especially in overcrowded countries that lack ‘legalized’ method of abortion, for those who
Can be seen as a way to reduce the need for clinical May be seen as a stimulus for pre-marital sex
abortions and unwanted pregnancies, which could lead and promiscuity, especially in teenagers and
to child neglect and depriving young parents of future young adults, due to the easy availability of
opportunities. condoms.
Allows either partner to maintain control of their Some methods may have physiological risks
bodies if the other wishes not to use birth control, such as birth control pills, or may cause
especially during pre-marital sex. infection if not done correctly, such as Filshie
clips.
PRE-NATAL CARE
Pre-natal or post-conceptual care refers to the behaviours or routines that a mother should adopt to
reduce the incidence of ill health or hindrance to development of the foetus. Before pregnancy, however,
the mother should ensure that she has the RUBELLA vaccine, as rubella can be fatal for the foetus. The
table below will summarize the main points of pre-natal care:
Avoiding foods that are precooked or May contain Listeria bacteria, which
unpasteurized milk infect foetus.
Rhesus If the mother is Rh-negative then she should be Without the anti-Rh antibodies, the
factor injected with anti-Rh antibodies if she has a Rh- mother’s own antibodies will attack
positive foetus. the Rh-positive foetus.
Smoking, one of the worst offenders of foetal malformation, will be discussed in the next page.
Keep in mind that whatever the mother takes into her bloodstream will transfer to the foetal bloodstream
via the chorion and umbilical cord connected to the placenta. Tobacco cigarettes contain a massive
number of chemicals that have detrimental effects to the human body. So we’ll look at the impact of three
of the main components: nicotine, carbon monoxide and tar.
In pregnancy, it is recommended that a healthcare provider check the health of the foetus. This is done by
checking the baby’s heart rate and other functions. The details may vary, but typical electronic fetal
monitoring may go like this:
• INTERNAL – This is usually done if the amnion has already ruptured and labour has begun. The
provider puts a small wire called a SCALP ELECTRODE through the cervix and attaches to the
baby’s scalp. The electrode is attached to a wire. The wire sends information about the foetal
heartbeat to a computer.