Biotechnology and Bioinformatics (1+1)

2.1.1. Introduction

The recombinant DNA technology is also referred to as gene cloning or molecular cloning. The discovery of restriction enzyme s and many other enzymes which can be useful as tools in gene manipulation have brought in vitro recombining of DNA to a reality and dependence on in vivo recombinational events came to an end.

Gene cloning may be aimed at getting more copies of a particular gene in desired host cell. In a way it is gene amplification achieved through gene cloning.

But it is also possible to have expression of gene in desired host cells after cloning. This will result into formation of product of that gene into new host cells. This can be useful for production of protein s into cells which are convenient to cultivate. Cloning can be done in bacterial host (e.g. E .coli) or in eukaryotes (e.g. yeast).

Basically cloning involves 4 steps.

1. The vector DNA is cleaved with one or more restriction enzymes.

2. The DNA to be clone d, the target or insert is joined to the vector, generating a recombinant molecule.

3. The recombinant DNA molecule is introduced into the host bacterial cell.

4. Transformed colony is selected and amplified.