BIOTECHNOLOGY QUESTIONS- Chapter 1

1. What is the basic difference between dNTP and ddNTP?

ddNTP differs from dNTP by having a H-tom attached at the 3-C rather than OH-
group and therefore ddNTP are unable to form a phosphodiester bond with the next
dNTP leading to chain termination.
2. ddNTP is important for DNA sequencing. Why?
Since they lack a OH-group, they lead to chain termination.
3. Automated DNA sequencing is an improvement of Sanger's method where
a. Fluorescently labelled ddNTPs are used for chain termination method
b. Fluorescently labelled dNTPs are used for chain termination method
c. ddNTPs are used for chain termination
d. PCR is used for making sequencing templates
4. What is the principle of blue-white screening?
Insertional inactivation of LacZ gene on the vector
5. Insertional inactivation is defined as
a. technique to identify desirable gene
b. technique to recognize non-recombinant cells
c. technique to recognize recombinant cells
d. technique to recognize specific cell types
6. In genetic engineering, the antibiotics are used
a. as sequences from where replication starts
b. to keep the cultures free of infection
c. as selectable markers
d. to select healthy vectors
7. What is the function of Mg2+ ions in PCR?
It is used to enhance the enzymatic activity of the process.
8. Why is ssDNA used in DNA sequencing?
It is used for sequencing of the DNA and so that the DNA can be read out.
9. How are fragments separated in RFLP?
Using Agarose gel electrophoresis
10. For whom does the RFLP pattern does not vary?
Identical twins
11. The major application of RFLP
DNA Fingerprinting
12. Use of PCR
Used for selective amplification of specific region of a DNA molecule.
13. PCR was invented by
Kerry Mullis (1985)
14. What is the source of Taq polymerase?
Thermus aquaticus
15. Which is the most imp step of PCR?
Denaturation
16.What is the process of binding of primer to the denatured stand called?
Annealing
17. Polymerase used for PCR is extracted from
Thermus Aquaticus
18. How many DNA duplexes are obtained from one DNA duplex after 4 cycles of PCR
16
19. Name the primers used for the PCR process
Single stranded DNA oligonucleotide
20. What is meant by one cycle in PCR
The geometric progression of DNA molecules from 2 to 4
21. E Coli cells divide every _____ minutes to clone large amounts of foreign DNA.
20
22. What does the protein made by Ecoli cells used for?
Therapeutics and other uses
23. What type of host cell can be used for proper folding and post translational modification?
Yeast
24. Which host cell cannot be processed in the presence of introns?
Ecoli
25. Yeasts serve as ____
large culture vessel
26. What kind of host cells can be grown in tissue cultures?
Plant and animal cells
27. Cells used to induce and manipulate to form whole organisms are _____
Plant and animal cells
28. Which type of gene transfer method that takes up DNA from surrounding and chemically
treated to become competent?
Transformation
29. What is the solution in which Ecoli becomes competent to take up DNA
Calcium chloride solution
30. Transformation is discovered by whom and when?
1970, Mendel and Higa
31. What are the charged substances that can be used for mixing of foreign DNA with host
cells?
Calcium phosphate, cationic lipase and DEAE dextran
32. What helps as natural genetic engineer?
Agrobacterium tumefaciens
33. Most commonly used vectors in cloning
plasmids & bacteriophages.
34. Features of plasmid-based vectors
plasmid extra chromosomal, self-replicating, circular etc.
35.Why Plasmid based vectors are ideal for cloning?
As it transfers traits on to host organism or versatile and easily manipulated.
36. expression vector –pSOMI and its function
production of somatostatin.
37. Earliest Plasmid
pBR322
38. Popular series of plasmid cloning vectors?
pUC family
39. During screening, what indicates the plasmid containing the insert?
The absence of Beta –galactosidase
40. Naturally occurring plasmid in plants?
Agrobacterium tumefaciens
41. LEU2 genocides for enzyme which is needed for synthesis of aa?
leucine
42. 2 phages that have been extensively modified for development for cloning vectors?
lambda and M13 phages.
43.What is lytic pathway?
44. Bacteriophages lambda has – DNA genome?
Double stranded, linear
45. ------- are important for packaging DNA into phage heads?
Cos sites or cohesive
46. Phage based vectors allow coning of DNA fragments upto?
23 kb in Size
48. M13 phage infects…?
E. coli having a pilus.
49. Restriction form replicates until there are about …………. copies in cell?
100
50. Major advantage of development vectors based on M13?
Genome less than 10kb in Size and can be manipulated exactly like plasmids.
51. Single stranded forms of cloned DNA are useful for various techniques …. and ….?
DNA sequencing, site directed mutagenesis.
52. Cosmids formed by combining features of …. and ….
Plasmids,cos sites of lambda phages.
53. Cosmids can clone DNA fragments upto…
45 kbp
54. Full Form of YACs?
Yeast Artificial Chromosomes.
55. YAC vectors contains sequence?
Telomeric.
56. Who developed the 1st rDNA molecule?
Stanley Cohen, Herbert Boyer, Paul Berg and Annie Cheng
57. Which tool of RDT is called molecular scissors?
Restriction Endonucleases (RE)
58. Who 1st discovered RE and won the nobel prize in 1978?
W. Arber, H. Smith and D. Nathan
59. Classify the RE and which is used in cloning experiments?
RE type 1, RE type 2 and RE type 3. RE type 2 is used in cloning experiments.
60. Write the microbial source and restriction sites of EcoR1 and Pst1.
EcoR1: Escherichia coli: 5’GAATTC3’
Pst1: Providencia stuartii: 5’CTGCAG3’.
61. How do sticky ends bind and generate a hybrid or recombinant DNA?
When 2 different DNA fragments containing EcoR1 recognition sites are cleaved and mixed
62. What is the defense mechanism of bacteria called?
Restriction-modification system.
63. Which types of cuts are termed as staggered?
Sticky ends.
64. What does the roman number at the end of a RE signify?
Order of discovery
___________________________________________________________________________