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  Recombinant DNA technology

Recombinant DNA technology:

It is a method in which the chosen DNA of one organism (Donor) is introduced to combine with the DNA of another organism termed as recipient organism. The result of it is that, the recipient organism acquires the genetic abilities of the donor. Altering the genome of an organism through introducing genes of interest is termed as gene manipulation or DNA recombinant technology. According to this mechanism has the capability to engineer new organisms, it is termed as genetic engineering.

Basic techniques of genetic engineering:

Bacterial cells comprise dissimilar types of enzymes. A number of these can cut DNA into fragments and others can join such type of fragments. For instance, restriction endonucleases that located in the year 1970 are involved in cutting DNA at particular sites. Therefore they are termed as molecular scissors. The enzyme DNA ligase discovered in the year 1966 works as a paste molecule to join DNA fragments. So the restriction endonuclease and the DNA ligase are the fundamental tools needed for genetic engineering.

The events of recombinant DNA (Deoxyribonucleic acid) technology are as follows:

1. The DNA of donor organism / gene of interest is isolated and cut into fragments through using restriction endonuclease.

2. They are connected to an appropriate replicon. Such type of replicon is termed as vector or cloning vehicle that is nothing but the extra chromosomal circular DNA that found in the cytoplasm of Eschrichia coli is termed as plasmid.  The plasmids are the most appropriate vectors.

3. The DNA of the vector is cut into fragments by using similar restriction endonucleases. Using the enzyme DNA ligase, the DNA fragments of donor and vector are attached together. This procedure is termed as splicing. The result of splicing hybrid DNA or recombinant DNA (rDNA) is acquired.

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4. The rDNA is present into the host cells like E.coli, Bacillus subtilis, Streptomyces sp. etc.

5. For this reason the host cells are treated along with the enzyme cellulase. Thus that the cell wall of host becomes permeable to the entry of rDNA.

The instructions of “foreign rDNA” are followed by the host organism. It carries on to multiply with the foreign DNA / gene of interest. After a very short time, this results in a colony of bacteria comprising rDNA fragments. Every colony is grown-up independently to acquire multiplication of rDNA fragments.  At the end we obtain some colonies having similar copies of rDNA fragments. This is termed as molecular coloning / gene coloning.

One time the gene for the production of human insulin from pancreatic cells is introduced into E.coli, the recipient cell generates human insulin. This is the method through which the human insulin is made to be generated by bacterial cell like E.coli.

 

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