Question

Steps of Recombinant DNA technology are given below:

A. Identification and isolation of the genetic material.

B. Fragmentation of DNA.

C. Obtaining the foreign gene product

D. Downstream processing.

E. Ligation of DNA fragmentation into the vector.

F. Isolation of desired DNA fragments.

G. Amplification of gene of interest.

H. Transfer of Recombinant DNA into the host cell/organism.

The correct sequence of steps is

1. A → D → C → B → E → G → F → H
2. A → B → F → G → E → H → C → D
3. H → F → G → E → A → D → B → C
4. C → A → B → D → F → E → G → H

Answer 1

Correct Answer - Option 2 : A → B → F → G → E → H → C → D

The correct answer is A → B → F → G → E → H → C → D.

• The complete process of recombinant DNA technology includes multiple steps, maintained in a specific sequence to generate the desired product
  • Identification and isolation of the genetic material: The first and the initial step in Recombinant DNA technology is to isolate the desired DNA in its pure form i.e. free from other macromolecules.
  • Fragmentation of DNA: The restriction enzymes play a major role in determining the location at which the desired gene is inserted into the vector genome. These reactions are called ‘restriction enzyme digestions’.
  • Isolation of desired DNA fragments.
  • Amplification of gene of interest: It is a process to amplify a single copy of DNA into thousands to millions of copies once the proper gene of interest has been cut using the restriction enzymes.
  • Ligation of DNA fragmentation into the vector: In this step of Ligation, joining of the two pieces – a cut fragment of DNA and the vector together with the help of the enzyme DNA ligase.
  • Transfer of Recombinant DNA into the host cell/organism: The recombinant DNA is introduced into a recipient host cell. This process is termed Transformation. Once after the insertion of the recombinant DNA into the host cell, it gets multiplied and is expressed in the form of the manufactured protein under optimal conditions.
  • Obtaining the foreign gene product: Multiplication/expression of the introduced gene in the host.
  • Downstream processing: Selection of the transformed host cells and identification of the clone containing the desired gene/DNA fragment.