BIOCHEMISTRY o Australia, Belgium, Canada, Denmark,

Human Genome Project Germany, Israel, Italy, Netherlands,

Dr. Guerrero Russia, Sweden, China
AFCG, REM, SMB
MAJOR INTERNATIONAL ORGANIZATIONS INVOLVED
 HUGO (Human Genome Organization)
GENOME  EC (European Council)
 Organism’s complete set of DNA  UNESCO
 DNA – paired strands
o Each strand is made of four nucleotide III. Deciphering the Human Genome
bases EXPERIMENTAL PROCEDURES
o Bases: Adenine (A), Thymine (T), a) Restriction Fragment Polymorphism (RFLP)
guanine (G) and cytosine (C) o Identification of sequence variations in
 A-T, C-G DNA sites that can be cleaved by
restriction enzymes
I. Informative Data b) Pulsed-field Gel Electrophoresis
HUMAN GENOME o Laboratory technique used by scientists
 Comprised of 23 pairs of chromosomes to produce DNA fingerprint for bacterial
o 22 autosomes and 1 sex chromosome isolates
 Smallest human chromosome: Y, 50M bp o PFGE Steps:
 Largest Human Chromosome: 1, 250M bp 1. Bacterial cells from an agar
 Karyotype: analysis of chromosomes via plate
microscope based on shape (size and banding 2. Mixes bacterial cells with
pattern) melted agarose and pours into
a plug mold
II. What is the Human Genome Project? 3. Bacterial cells are lysed so DNA
GOALS is free
 Identify the approximate 100,000 genes in 4. Loads DNA gelatin plug into a
human DNA gel and into an electric field
 Determine the sequences of the more than 3 B that separates DNA fragments
bases that make up human DNA 5. Stained so DNA can be seen
 Store information in database under UV light
 Develop tools for data analysis c) PCR (Polymerase Chain Reaction)
 Address ethical, legal and social issues that o In vitro amplification of specific nucleic
arise from genome research acids
HUMAN GENOME PROJECT o PCR Markers
 International effort to determine the sequence  Based on short, repetitive DNA
of the human genome and identify the genes sequences widely distributed in
that it contains the human genome
 Ran from 1990 - 2003 a) Yeast Artificial Chromosome (YAC)
 US government project coordinated by DOE o Genetically engineered chromosomes
(Department of Energy) and NIH (National derived from the DNA of yeast
Institutes of Health) o To isolate and propagate very large
 Budget: 3 billion dollars segments of DNA in a yeast host
 15-year effort b) Sequence Tagged Site
 International consortium o Short unique DNA sequence that can be
o US, France, UK, Japan
amplified by PCR (easily detected)

Human Genome Project | Bacolor, Carag, Miguel |1

 c) Positional Cloning o Sequence variations in DNA sites that
o Markers are used for gene hunts can be cleaved by restriction enzymes
o Once the gene is located, physical maps  Short Tandem Repeat Polymorphism (STRP)
are used to obtained flanking DNA o Advantages:
segments for further detailed study  Repeated up to thousands of
(mostly pertains to regulation of gene times throughout the genome
function)  Even distribution throughout
o Identifying gene inflicting a disease the genome
(inheritable disease)  Amplifiable by PCR
 Number of repeats vary among
IV. Mapping and Sequencing the Human Genome individuals
MAPPING
 Divide each chromosome into small segments
 Arranging them sequentially on the V. Mapping Methods
chromosome MACRO-RESTRICTION MAPS
GENETIC MAP a) Top-down mapping:
 Depicts the order by which genes are arranged o Fragmenting chromosomes with a rare
along a chromosome restriction enzyme into large pieces ->
 Sequence (genetic map) is facilitated by known ordered -> subdivided -> mapped
markers: genes or other DNA stretches o Smaller pieced mapped together
 Distances between markers are measured in o Result: more continuity and less gap
centimorgans (cM) than the contig method, but it has a
o Unit of measure of recombination lower map resolution
frequency b) Contig Map (Bottom-up mapping)
o 1 genetic cM is about 1 M base pairs o Cutting a chromosome into small
(bp) (on physical distance) pieces, each cloned and ordered,
 Assisted in chromosomal location of several forming contiguous DNA blocks
inherited diseases (Sickle Cell disease, cystic c) STS-Content Mapping
fibrosis, Tay-Sachs disease, Fragile X syndrome, o Provides the means to establish these
Myotonic dystrophy, ataxia telangiectasia) overlaps between each clone and its
GENE LINKAGE MAP nearest neighbors
 Shows the relative location of a specific DNA o If 2 clones share even a single STS, they
marker along the chromosome can reliably be assumed to overlap.
 Constructed by observing how frequently two d) Radiation Hybrid Mapping
markers are ‘inherited’ together o Involves fragmentation of
 The closer the markers are to one another on chromosomes in cultured cells with high
the same chromosome, the more tightly linked doses of X-rays -> Incorporation of
they are, the more likely they will be passed to fragments into stable cell lines
the next generation VI. Terminologies
PHYSICAL MAP CONTIG (CONTIGUOUS)
 Shows actual sites of genes on the genome  Set of overlapping DNA segments that together
 Comprised of landmarks, (restriction enzymes represent a consensus region of DNA
and STS) providing reference points relative to  Organized set of DNA clones that collectively
which DNA sequence such as genes can be provide redundant cloned coverage of a region
localized too long to clone in one piece
 Restriction Fragment Length Polymorphism COSMID
(RFLP)  Often used as a cloning vector in genetic
engineering
Human Genome Project | Bacolor, Carag, Miguel |2
  Can be used to build genomic libraries  determine pedigree for seed or livestock breeds
 Designed for cloning fragments of DNA (20k-40K AGRICULTURE AND LIVESTOCK
base pairs)  disease, insect, and drought-resistant crops
FISH (FLUORESCENCE IN SITU HYBRIDIZATION)  healthier, more productive, disease-resistant
 Physical mapping technique employing farm animals
fluorescein-labelled DNA probes that can detect  more nutritious produce
segments of the human genome by DNA  bio-pesticides
GEN BANK  edible vaccines incorporated into food products
 Public database of DNA sequence operated by  new environmental cleanup uses for plants like
NIH tobacco
 Accessible freely and without restrictions to all EVOLUTION AND HUMAN MIGRATION
scientists in industry and academe  study migration of different population groups
based on female genetic inheritance
I. Gene Therapy  study mutations on the evolutionarily stable Y
 The prime benefit to be derived from the Human chromosome to trace lineage and migration
Genome Project, in which  compare breakpoints in the evolution of
 Defective genes inflicting congenital diseases are mutations with ages of populations and historical
replaced by functional genes events
 Genes are likely to be the key targets for new types RISK ASSESSMENT
of therapy (eg. if a gene is mutated so it is  assess health damage and risks caused by
permanently active, gene therapy could inactivate radiation exposure, including low-dose exposures
the protein, or if a gene has been turned back on or  assess health damage and risks caused by
a second, normal copy is introduced) exposure to mutagenic chemicals and cancer-
causing toxins
II. Benefits Of Human Genome Project  reduce the likelihood of heritable mutations
MEDICAL BENEFITS
 Improved diagnosis of disease III. More Information (FYIs)
 Earlier detection of predispositions to disease  The Human Genome Project assembles 12,000
 Rational drugs design bases every minute
 Gene therapy and control systems for drugs  15 billion raw base pairs were sequenced to
 Pharmacogenomics “personal drugs” reach the two billion milestone
 Organ replacement  Each area of a chromosome is sequenced at least
MICROBIAL GENOME RESEARCH four to five times to ensure that the data
 New energy sources (biofuels) deposited into the GenBank is accurate
 Environmental monitoring to detect pollutants  2 billion of the 3 billion “letters” that constitute
 Protection from biological and chemical warfare the genetic instruction book of humans have
 Safe, efficient toxic waste clean up been deciphered and deposited into GenBank
 As per 2003’s technology, the Human Genome
Project is as complete as it can be.
DNA FORENSICS  Small gaps that are unrecoverable in any current
 Identify potential suspects at crime scenes sequencing method remain.
 exonerate wrongly accused persons  New technologies will have to be invented to
 identify crime and catastrophe victims obtain the sequence of these regions.
 establish paternity and other family relations  Even though the Human Genome Project has
 detect bacteria and other organisms that may been completed, scientists continue to develop
pollute air, water, soil, and food and apply new technologies to the few
 match organ donors with recipients in transplant remaining refractory problems.
programs

Human Genome Project | Bacolor, Carag, Miguel |3

  Continue to support a wide range of research to
develop new sequencing technologies, to
interpret the human sequence and to use the
newfound understanding of the human genome
to improve human health
 About 100 cancer genes have been already been
found, most of which are from rare leukemia and
lymphomas (<10% of all human cancer)
 Common adult cancers (breast, colon, prostate,
lung and ovary) which account for 80% of the
cancer burden -> only about 30 genes are known.

 REFERENCES
 PowerPoint presentation of Dra. Charisse
Guerrero
 Human Genome Project FAQs

Human Genome Project | Bacolor, Carag, Miguel |4