Question

How do you insert a gene into a plasmid?

Answer 1

Hint: The insertion of a gene into the plasmid is known as gene cloning, which takes place to insert a specific gene into the plasmid vector for its transformation. The insertion takes place with the help of restriction enzymes and DNA ligase.

Complete answer:
In biotechnology, the insertion of a gene in a vector plasmid aims to develop a new genetic combination that takes place in laboratory conditions by which the host cell multiplies. The insertion starts with the use of restriction enzymes which is also known as restriction endonuclease which cut the gene and circular plasmid at a specific point through the sugar-phosphate backbone, both the gene and plasmid produce compatible ends. Then the cut fragment of both gene and plasmid gets purified and ligated with the help of ligase. For cutting of DNA and plasmid, the sequence of both must be the same, and this type of sequence is called a palindromic sequence. Restriction endonuclease is of two types, one which produces a short single-stranded end which joins with the single-stranded end of a specific sequence whereas the second endonuclease cuts both the strands of DNA at the same recognition site. After the insertion of a gene in the plasmid, the new plasmid is introduced in the host where it multiplies and produces multiple copies of the plasmid which help to induce new genes.

Note:
Restriction endonuclease is also a nuclease enzyme. The nuclease is divided into exonuclease and endonuclease. The exonuclease helps in the cleavage of DNA molecules from the end, whereas endonuclease cuts DNA from inside at its specific site which is also known as recognition sequence.