Question

(a) Mention the role of vectors in recombinant DNA technology. Give any two examples
(b) With the help of diagrammatic representation only, show the steps of recombinant DNA technology.

Answer 1

Hint: Recombinant DNA technology is the part of molecular biology, which deals with the expression of the gene of interest in a certain host mainly E. coli, and extracting the desired protein from the host cells. There are several components present in this technique like restriction enzymes, vectors, the gene of interest, host cell, etc.

Complete answer:
(a) In recombinant DNA technology, there are various components one of them is the vector, it is the most important component of recombinant DNA technology. Vectors are the molecular carriers that carry the gene of interest to the host cells. Vectors can replicate independently without the control of chromosomal DNA. In this method, a foreign short piece of DNA which is mainly the gene of interest is been inserted to the vector. Then after doing this, this vector is being transformed into the bacterial cell host mainly E. coli cells, so that it can replicate inside the host cells and after that protein of interest can be isolated from it. The two examples of the vector can be a bacteriophage vector and a plasmid vector.

(b)

- There are various steps discussed below for the Recombinant DNA technology.
- Isolation of genetic material in which rDNA free from other macromolecules gets isolated in pure form.
- Restriction Enzyme digestion helps in the cutting of DNA at specific locations.
- Amplification using PCR (Polymerase Chain Reaction) is a method in which the formation of multiple DNA copies takes place by using the enzyme DNA polymerase.
- Ligation of DNA molecules helps in joining the two pieces of DNA by using a DNA ligase enzyme.
- Insertion of recombinant DNA into the host
- Isolation of recombinant cells

Note: Recombinant DNA technology is a very famous molecular biology technique, which has several steps to get the desired protein from the gene of interest. It first involves getting the desired gene of interest, then cutting the vector with the restriction enzymes then ligating the gene of interest to the vector and then transforming it to the host cells like E. coli and there the vector will replicate and form desired proteins, which will be isolated.