Biotechnology: Principle & Processes

1. Name the technique used for amplification of DNA? [1]

2. Name the enzyme responsible for removal of 5 – phosphate group from nucleic acid?
[1]
3. Which enzymes are known as “molecular Scissors”? [1]
4. Why does DNA moves towards anode in gel electrophoresis. [1]
5. Name the commonly used vector for transformation in plant cell? [1]
6. Differentiate between plasmid DNA and chromosomal DNA? [2]
7. What is the role of enzyme “Ligase” in genetic Engineering? [2]
8. What is Bioreactor? What are the advantages of Stirred tank. Show diagrammatically a
simple Stirred tank Bioreactor? [2]
9. What is “Insertional Inactivation”? [2]
10. What are Restriction enzyme? Why do bacteria have these restriction enzymes?
Show diagrammatically a restriction enzyme its recognition & the product it
produces. [2]
11. What are the two basic techniques involved in modern Biotechnology? [2]
12. Represent diagrammatically the E. coli. Cloning vector PBR 322. [2]
13. Mention the important tools required for genetic engineering technology? [3]
14. What are the two basic techniques involved in modern Biotechnology? [3]
15. Expand PCR? Describe the different Steps involved in this technique? [5]
 ANSWER
1. Polymerase Chain Reaction (PCR).
2. Phosphatases.
3. Restriction Endonucleases.
4. The phosphate backbone of the DNA molecule is negatively charged, therefore when
placed in an electric field, DNA fragments will migrate to the positively charged
anode.
5. Agrobacterium tumefaciens.
6.
PLASMID DNA CHROMOSOMAL DNA
It is always double stranded. It may be single stranded or double stranded.
It is circular. It is linear or circular.
It is naked without histone protein. It is coated with histone protein.
It does not carry any vital gene necessary for It carries vital genes necessary for cell.
cell.
It can replicate independent of main It replicates with genome.
genome.
It does not act as genetic factor. It acts as genetic factor.
Introns are absent. Both exons and introns are present

7. DNA ligase enzyme is used to join two DNA fragments from their ends. It facilitates
the joining of DNA strands together by catalyzing the formation of a phosphodiester
bond.
8. Bioreactors are the large volume (100-1000 L) vessels in which raw materials are
biologically converted into specific products, individual enzymes, etc, using
microbial plant, animal or human cells.
The stirred-tank reactor is usually cylindrical or with a curved base to facilitate the
mixing of the reactor contents. The stirrer facilitates mixing and oxygen availability
throughout the bioreactor.
9. The insertional activation of alpha-galactosidase enzyme, i.e. by inserting the
desired gene in the coding region of enzyme, results in inactivation of alpha-
galactosidase gene in recombinants. The recombinant on transformed hosts are
unable to produce any colour when grown on chromogenic substrate, thus acting as
a selectable marker to differentiate recombinants from non-recombinants.
10. Restriction enzyme, also called restriction endonuclease, a protein produced by
bacteria that cleaves DNA at specific sites along the molecule. In the bacterial cell,
restriction enzymes cleave foreign DNA, thus eliminating infecting organisms.

11.Principles of biotechnology are based on the concept of the following

techniques:
(i) Genetic engineering is the technique to alter the chemistry of genetic material
(DNA/RNA), to introduce these into another organisms and thus, change the
phenotype of the host organism.
(ii) Adequate maintenance of sterile conditions to support growth of only the
desired microbes/eukaryotic cells in large quantities for the manufacture of
biotechnological products like antibiotics, vaccines, enzymes, etc.
 12.

13. (i) Restriction enzymes (ii) Polymerase enzymes

(iii) Ligases (iv) Vectors
(v) Competent host organism.
14. Same as question no 11
15.Polymerase Chain Reaction (PCR)
Steps in PCR
i) Denaturation of double stranded DNA is carried out by applying high temperature
of 95°C for 15 seconds. Each separated single strand acts as a template for DNA
synthesis.
ii) Annealing is carried out by two sets of primers, which are added in the reaction.
They anneal to the 3′end of each separated strand. Primers act as initiator of
replication.
iii) Extension is done by DNA polymerase by adding nucleotides complementary to the
template in the reaction.