RECOMBINANT DNA TECHNOLOGY

This field of biotechnology encompases a number of methodologies that enable new combination of genetics material to be artificially construted in laboratory.

In other words

Recombinant DNA technology involves using enzymes and various laboratory techniques to manipulate and isolate DNA segments of interest. This method can be used to combine DNA from different species or to create genes with new functions. The resulting copies are often referred to as recombinant DNA.

rDNA

Recombinant DNA is a form of the artificial DNA that is made through the combination or insertion of one or more DNA strands, therefore combining DNA sequences as per your requirement.

DICOVERY

·       Discovery of DNA structure- 1953 watson and crick.

·       Isolation of DNA ligase- 1967 (for the formation of phosphodiester bond with the help of ATP).

·       Isolation of restriction endonucleases around- 1970.

·       Formation of recombinant DNA – 1972.

·       First human protein made by using rDNA technology- Somatostatin in E.coli.

·       First plasmid vector capable of replicate within host 1973.

RECOMBINANT DNA TECHNOLOGY GOALS

·       To isolate and characterize a gene.

·       To make desired and alternation in one or more isolated gene.

·       Transfer transgene into living cells.

·       Artificially synthesize new gene.

·       Understanding the heriditory disease & the cure.

·       Improving human genome.

 

 

 

 

 

 

 

 

 

 

Steps involved in rDNA experiment-

 

Identification of desired gene

 

 

 

 

Isolation of desired gene with the help of restriction enzyme

 

 

 

 

The vector cut with theb same restriction enzyme

Vector and isolated gene is ligated together with help of DNA

Amplification and transfer of desired gene into the progeny

The recombinant vector is introduce into the host cell

 

 

 

 

 

 

 

 

 

 

 

 

 

 

                         

 

                                          

 

 

 

 

 

 

 

Gene cloning

The basic steps in a gene cloning experiment are follows.

 

 

1.      A fragment of DNA containing the gene to be cloned, is inserted into a circular DNA molecule called vector, to produce a chimera or recombinant DNA molecule.

2.      The vector act as  vehicle that transport  the gene into a host cell, in which is usually a bacterium, although other types of living can be used.

3.      Within the host cell