• PRINCIPLES & TOOLS OF BIOTECHNOLOGY

1. EFB full form -

2. The 2 core technique that enable the birth of modern biotechnology are -
3. ________ is a specific DNA sequence which is responsible for initiating replication. (NEET)
4. Autonomously replicating circular extra-chromosomal DNA is _______ (NEET)
5. First recombinant DNA was formed by ______ and _______ (scientists) in the year _____, by
working on _________ bacteria. (NEET)
6. Plasmid of _____ bacteria was taken and then it was inserted after modification in _____ bacteria.
7. ________ are known as molecular scissors. (NEET)
8. The linking of antibiotic resistance gene with the plasmid vector became possible with the enzyme
________ (NEET)
9. 3 basic steps in genetically modifying as organism is -
10. In 1963, the two enzymes responsible for restricting the growth of bacteriophage in
Escherichia coli were isolated. What was the function of both of them ?
11. First restriction endonuclease was - (NEET)
12. What is the difference between endonuclease and exonuclease ?
13. How are restriction endonucleases named ?
14. Hind II palindromic sequence has ___ no. of base pairs.
15. Today we know more than ____ (no.) restriction enzymes that are isolated from ____
(no.) strains of bacteria.
16. In EcoRI, R is derived from the name of ______ (NEET)
17. EcoRI comes from bacteria - (full name) (NEET)
18. Each restriction endonuclease recognizes a specific _________ in the DNA. (NEET)
19. Palindromic nucleotide sequence of EcoRI is -
20. The DNA fragments are separated by a technique known as - (NEET)
21. In gel electrophoresis, the matrix used is of ______ (material) which is a natural polymer extracted
from _______ (NEET)
22. Agarose gel provides ______ effect. (NEET)
23. DNA fragments are visualised only after staining it with _______ followed by exposure to _______
24. Red/orange coloured bands of DNA are seen.
25. The separated bands of DNA are cut out from the agarose gel and extracted from the gel piece. This
step is known as ________
26. All types of plasmids are present in equal numbers
in the cell. T/F
27. ________ is responsible for controlling copying numbers of the linked DNA. (NEET)
28. The normal E coli. carry resistance to ampicillin. T/F (NEET) A B
H
29. Give examples of selectable markers for E. coli (4) -
30. Cloning sites should be preferably single/double. G
31. What are transformants? C
F
32. What are recombinants? P Q
33. The two antibiotic resistance genes in pBR322 are -
DigaQ. 1 D
34. pBR322 have restriction sites for (6) -
35. Rop gene codes for - R
36. Restriction sites in tetR are - (2) S
37. Restriction sites in ampR are - (2)
38. Restriction site in rop is -
39. "Insertional inactivation" help is selection of transformants/recombinants. E
40. In chromogenic selection, DNA is inserted in the coding sequence of _______ enzyme. (NEET)
41. In absence of any insert, the colonies give ______ colour.
42. ________ is able to deliver a piece of T-DNA. (NEET)
43. Agrobacterium tumefaciens is a pathogen of monocot/dicot plants.
44. T-DNA transforms normal cells to tumor cells. T/F
45. ______ in animals have the ability to transform normal cells to cancerous cells.
46. Plasmid of Agrobacterium tumefaciens is called _______
47. Host are made competent by treating them with specific concentration of _____ion.
48. Heat shock is of ____ °C.
49. Recombinant DNA is forced into cells by changing temperature. Tell how ?
50. In ________ (method), recombinant DNA is directly injected into the nucleus of an animal cell.
51. In plants, cells are bombarded with high/low velocity microparticles of ____ and ____ coated with
DNA in a method known as _____ or _____

DigaQ. 2. What process is going on?

 • PROCESSES OF RECOMBINANT DNA TECHNOLOGY

52. _______ is used to break bacterial membranes, ______ is used to break plant cells and
______ is used to break fungal cells. (NEET)
53. Purified DNA ultimately precipitates out after the addition of _________ (NEET)
54. Why does DNA precipitate after adding chilled ethanol ?
55. _________ is employed to check the progression of a DigaQ. 3
restriction enzyme digestion. (NEET)
56. DNA moves towards the positive electrode (cathode). T/F G
57. PCR full form - (NEET) A
58. The 3 steps of PCR are - F
59. If a PCR cycle runs 30 times, it will produce _____ times B
the initial amount of desired DNA sequence put into the system. E
60. ______ no. of copies of desired genes will be produced after
D
the end of first 5 cycles of PCR.
61. DNA polymerase used in PCR is isolated from bacteria - (NEET)
62. The DNA polymerase in PCR is known as _______
63. If any protein encoding gene is expressed in a heterologous C
host, it is called ________ protein.
64. In continuous culture systems, cells maintain themselves in DigaQ. 4
log phase. T/F
65. ________ phase in the physiologically most active phase. A
66. In bioreactors, ____-____ litres of culture can be processed.
B
67. The commonly used bioreactors are of ______ type.
68. A stirred-tank has a curved/flat base.
69. The bioreactors have many systems attached to them.
Name them all. (6)
70. ______ and ______ are collectively referred to as
downstream processing. (NEET)
71. The downstream processing and quality control testing
vary from product to product. T/F
• PRINCIPLES & TOOLS 26. F
1. European Federation of Biotechnology (EFB) 27. Ori
2. Genetic engineering & Bioprocess Engineering 28. F
3. Origin of replication 29. Ampicillin, chloramphenicol, tetracycline or
4. Plasmid kanamycin resistant genes
5. Stanley Cohen and Herbert Boyer, 1972, 30. Single
Salmonella typhimurium 31. Transformants are the cell that has taken the
6. Salmonella typhimurium, E coli. additional genetic material (it may be natural or
7. Restriction endonuclease genetically engineered)
8. DNA ligase 32. Recombinants are the cells who have taken up
9. Identification of DNA, introduction of DNA, the genetically engineered genetic material.
maintenance of introduced DNA 33. Ampicillin and tetracycline
10. One added methyl groups to DNA, while the 34. Hind III, EcoR I, BamH I, Sal I, Pvu II, Pst I, Cla I
other cut DNA 35. Proteins involved in replication of plasmid
11. Hind II 36. BamH I, Sal I
12. Exonucleases remove nucleotides from the ends 37. Pvu I, Pst I
of the DNA whereas, endonucleases make cuts at 38. Pvu II
specific positions within the DNA 39. Recombinants
13. First letter comes from genus and second two 40. β-galactosidase
letters come from species, then one letter from the 41. Blue
strain and the Roman numbers following the names 42. Agrobacterium tumefaciens
indicate the order in which the enzymes were 43. Dicot
isolated from that strain of bacteria. 44. T
14. 6 45. Retrovirus
15. 900, 230 46. Ti Plasmid
16. Strain 47. Calcium
17. Escherichia coli RY 13 48. 42°C
18. Palindromic nucleotide sequence 49. First cells are incubated on ice, then heat shock
19. GAATTC is given, and then again incubated on ice
20. Gel electrophoresis 50. Micro-injection
21. Agarose, sea weeds 51. High, gold and tungsten, biolistics or gene gun
22. Sieving
23. Ethidium bromide, UV radiation
24. Orange
25. Elution
• PROCESSES OF RECOMBINANT IV. Temperature control system
DNA TECHNOLOGY V. pH control system
52. Lysozyme, cellulase and chitinase VI. Sampling ports
53. Ethanol 70. Separation and purification
54. Ethanol has a low dielectric constant. As DNA is 71. T
a polar molecule, it would be more soluble in polar • DigaQs
solvents like water. As ethanol is less polar, it DigaQ. 1 – pBR322
precipitates out in it. A – Cla I P – ampʳ
55. Agarose gel electrophoresis B – Hind III Q – tetʳ
56. F, anode C – BamH I R – ori
57. Polymerase chain reaction D – Sal I S – rop
58. Denaturation, annealing and extension E – Pvu II
59. 2³⁰ F – Pst I
60. 6 (you may think that ans should be 2⁵ = 32, G – Pvu I
but on close observation you will notice that desired H – EcoR I
DNA sequence do not start to form till the 3rd DigaQ. 2 - Spooling
step) DigaQ. 3 - Simple stirred-tank bioreactor
61. Thermus aquaticus A – Acid/base for pH control
62. Taq polymerase B – Steam for sterilization
63. Recombinant C – Sterile air
64. T D – Culture broth
65. log/exponential E – Flat bladed impeller
66. 100-1000 F – Foam breaker
67. Stirring type G – Motor
68. Curved DigaQ. 4 - Sparged stirred-tank bioreactor through
69. Systems in bioreactors which sterile air bubbles are sparged
I. Agitator system A – Increased surface area for oxygen transfer
II. Oxygen delivery system B – Gas entrainment
III. Foam control system