What does 'Hind' and 'III' refer to in the enzyme 'Hind III'?

1

(a)

The first letter 'H' indicates the genus of the organism, from which the enzyme was isolaled, i.e. H-genus, Haemophilus.
The roman number (IIl) denotes the sequence in which the restriction endonuclease from that particular genus, species and strain of bacteria have been isolated, i.e. third restriction endonuclease to be isolated from this species.

How is copy number of the plasmid vector related to yield of recombinant protein?

1

(a)

The recombinant DNA can multiply as many times as the copy number of the plasmid vector. thereby determning the yield of recombinant protein So, higher the copy number of the plasmid vector, higher will be the yield of recombinant protein.

Would you choose an exonuclease for producing a recombinant DNA molecule?

1

(a)

No, because exonucleases will remove the nucleotides from the ends DNA molecule and not at any particular desired region.

What does 'competent' refer to, in competent cells used in transformation experiment?

1

(a)

Competent refers to the ability of the host cell to take up tha alien or recombinant DNA

What is the significance of adding proteases at the time of isolation of genetic material (DNA)?

1

(a)

Proteases remove the proteins associated with the DNA.

Name a recombinant vaccine that is currently being used in vaccination pro-grammes

1

(a)

Hepatitis-B vaccine.

What modification is done on the Ti-plasmid of Agrobacterium tumefaciens to convert it into a cloning vector?

1

(a)

Tumour inducing (Ti) plasmid of Agrobacterium tumefaciens is modified into a cloning vector by removing/ altering the gene responsible for its pathogenic property.

What is the role of CaCl₂ in the preparation of competent cells?

1

(a)

CaCl₂ increases the efficiency of the competent cells to pick up DNA  through the pores in its cell wall

Restriction enzymes present in the cloning site of a vector should not have more than one recognition site. Comment.

1

(a)

If the restriction enzymes have more than one recognition site in a vector, than the vector itself will get fragmented on treatement with the restriction enzyme.

Name the enzyme responsible for cleavage in the following sequence:
               \(\downarrow \)  
5' ___ G A A T T C ____3'
3' ___C T T A A G ____5'
                             \(\uparrow \)

1

(a)

Type II restriction enzyme obtained from Escherichia coli, called EcoRI.