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Practice each question, then open the answer or worked solution.
How is the copy number of the plasmid vector related to the yield of the recombinant protein?
Ans: Higher number of plasmid vectors results in the yield of a higher number of recombinant proteins.
Would you choose an exonuclease while producing a recombinant DNA molecule?
Ans: No, I would not choose exonuclease while producing a recombinant DNA molecule because it removes nucleotides from the ends of the DNA, and thus it cannot help in producing circular DNA.
What does H in ‘d’ and ‘III’ refer to in the enzyme HindIII?
Ans: No, I would not choose exonuclease while producing a recombinant DNA molecule because it removes nucleotides from the ends of the DNA, and thus it cannot help in producing circular DNA.
Restriction enzymes should not have more than one site of action in the cloning site of a vector. Comment.
Ans: Restriction enzymes should not have more than one site of action in the cloning site of a vector because the presence of more than one recognition site will create many fragments of DNA which will complicate the process.
What does ‘competent’ refer to in competent cells used in transformation experiments?
Ans: DNA is a hydrophilic molecule and so it cannot pass through the cell membrane thus the cell is made competent (pores are created) by treating suitable divalent ions. As a result, bacterial cells take up the DNA through the pores in the cell wall.
What is the significance of adding proteases at the time of isolation of genetic material (DNA).
Ans: The significance of adding proteases is that it helps in removing protein during the process of obtaining pure DNA
While doing a PCR, ‘denaturation’ step is missed. What will be its effect on the process?
Ans: During PCR, if denaturation is missed then primers will not join the template and this will halt the further process i.e., no extension, no amplification and at the end, large numbers of DNA cannot be made.
Name a recombinant vaccine that is currently being used in a vaccination program.
Ans: Hepatitis B vaccine is currently being used in a vaccination program.
Do biomolecules (DNA, protein) exhibit biological activity in anhydrous conditions?
Ans: Biomolecules such as DNA, protein do not exhibit biological activity in anhydrous conditions as they may get damaged under anhydrous conditions and proteins get denatured.
What modification is done on the Ti plasmid of Agrobacterium tumefaciens to convert it into a cloning vector?
Ans: Agrobacterium tumefaciens has the ability to infect plant cells and then transfer a DNA sequence (Ti plasmid) and cause a crown gall disease.
This plasmid vector is disarmed and then used as a cloning vector for delivering genes of interest to plants and animals.