Question

Enlist the basic steps involved in recombinant DNA technology.

Answer 1

Recombinant DNA is the DNA that is composed of sequences that are derived from different sources. The process of recombination DNA technology consists of the following steps:

1. Isolation of genetic material (DNA)

DNA is enclosed within the membrane. So, to extract it out, the cell wall is broken down to release the genetic material. This can be done by using cell wall digesting enzymes such as Lysozymes for bacterial cells, cellulose for plant cells and chitinase for fungi cells.

2. Cutting of DNA at specific locations

This is done by using restriction enzymes that cut DNA at specific recognition sequences and give the desired DNA fragments.

3. Joining of DNA fragment

The DNA fragments generated with a cloning vector can be done by sticky ends ligation, blunt-end ligation and homopolymer tailing.

4. Insertion of DNA into the host cell.

The recombinant DNA can be inserted into the host cell by transformation, transduction, and vectorless gene transfer.

5. Selection and screening of transformed cells

The selection and screening of transformed cells can be by any of the following methods- immunological method, nucleic acid hybridization and blue-white screening or insertional inactivation.