How to get a gene of interest?

GENE CLONING:

"The cloning in which identical copies of genes are produced is called gene cloning".

PCR (Polymerase Chain Reaction):

"The reaction which is used for production of lesser number of gene copies within test tube".

While, in case of formation of large gene copies recombinant DNA technology is used.

RECOMBINANT DNA TECHNOLOGY:

Recombinant DNA means a DNA with two different combination of genetic materials. (It is also called Chimeric DNA).

Method for Production of Recombinant DNA:

(i) Interest: Gene of interest (OR) selection, which is used to be cloned.

(ii) Cut out: Scissors enzymes (restriction endonuclease) to cut out the gene of interest.

(iii) Placement: Molecular carrier or VECTOR, on which gene of interest could be placed

(iv) Introduction: The gene of interest along with the vector is then introduced into an expression system, as a result of which a specific product is made.

Three ways to get the gene of interest:

(i) Isolation of gene from the chromosomes.

(ii) Synthesis of gene chemically.

(iii) Making of gene from mRNA.

Procedure:

(i) Isolation by restriction enzyme: The gene of interest can be isolated from the chromosomes by cutting restriction endonuclease is used to cut on the flanking sites of the gene.

(ii) Synthesis of small genes: In case of small genes they can also be synthesized in the laboratory.

(iii) Use of reverse transcriptase: Genes may be synthesized from mRNA by using the reverse transcriptase (an enzyme). This kind of DNA is called complementary DNA i.e, cDNA