Question

Explain the steps involved in gene cloning.

Answer 1

Answer:

(1) Isolation of DNA (Gene of Inter­est) Fragments to be Cloned

(2)Insertion of Isolated DNA into the a Suitable Vector to Form the Recombinant DNA

(3)Introduction of the Recombinant DNA into a Suitable Organism known as

Host

(4)Selection of the Transformed Host Cells and Identification of the Clone Con­taining the Gene of Interest

(5)Multiplication/Expression of the Introduced Gene in the Host

(6)Isolation of the Multiplied Gene Copies/Protein Expressed by the Intro­duced Gene

(7)Purification of the Isolated Gene Copy/Protein

Answer 2

Answer:

Gene cloning involves four steps: isolation, ligation, transformation, and selection.

1. In isolation, an enzyme is used to break DNA at a specific base sequence. This is done to isolate a gene.

2. During ligation, the enzyme DNA ligase combines the isolated gene with plasmid DNA from bacteria. (Plasmid DNA is circular DNA that is not part of a chromosome and can replicate independently.) The DNA that results is called recombinant DNA.

3. In transformation, the recombinant DNA is inserted into a living cell, usually a bacterial cell. Changing an organism in this way is also called genetic engineering.

4. Selection involves growing transformed bacteria to make sure they have the recombinant DNA. This is a necessary step because transformation is not always successful. Only bacteria that contain the recombinant DNA are selected for further use.

Explanation: