13.  Discuss the major steps involved in gene cloning.
State the basic steps involved in genetical modification of an organism.

Dear Student,

The steps involved in the process of cloning of gene in vectors are as under:

1. Preparation of cloning vector: In this step, the natural vectors like plasmids are obtained from the various bacteria or vectors are artificially produced by using genetic engineering techniques.

2. Isolation of foreign gene (desired gene): The desired gene is isolated from the cells of an organism by treating the cells with various enzymes like lysozymes, proteases, ribonucleases, etc.

3. Insertion of an isolated desired gene into a vector: DNA is cut into fragments with the help of restriction enzymes. By the gel electrophoresis technique, the DNA fragments obtained after restriction are isolated. The hybridization of 'gene of interest' with vector is done to obtain recombinant DNA. This step requires the enzyme DNA ligase.

4. Insertion of a vector in the host cell: This is done by the help of various methods like transformation in bacteria. 

5. Cloning of the desired gene takes place inside the host cell.

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