Mention the methods of making bacteria capable to take a rDNA.
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Answers
Explanation:
1.Isolation of Genetic Material
We already know that the genetic material of all living organisms is ‘nucleic acid’. In most organisms, it is DNA, whereas in some it is RNA. The first step in rDNA technology is to isolate the desired DNA in its pure form i.e. free from other macromolecules.
2.Restriction Enzyme Digestion
Restriction enzymes act as molecular scissors that cut DNA at specific locations. These reactions are called ‘restriction enzyme digestions’. They involve the incubation of the purified DNA with the selected restriction enzyme, at conditions optimal for that specific enzyme.
3.. Amplification Using PCR
Polymerase Chain Reaction or PCR is a method of making multiple copies of a DNA sequence using the enzyme – DNA polymerase. It helps to amplify a single copy or a few copies of DNA into thousands to millions of copies. PCR reactions are run on ‘thermal cyclers’ using the following components:
Template – DNA to be amplified
Primers – small, chemically synthesized oligonucleotides that are complementary to a region of the DNA.
Enzyme – DNA polymerase
Nucleotides – needed to extend the primers by the enzyme
4.Insertion of Recombinant DNA Into Host
In this step, the recombinant DNA is introduced into a recipient host cell. This process is ‘Transformation’. Bacterial cells do not accept foreign DNA easily. Therefore, they are treated to make them ‘competent’ to accept new DNA. (The topic – Tools of Biotechnology explains a few ways to make cells competent..
.5. Obtaining Foreign Gene Product
The recombinant DNA multiplies in the host and is expressed as a protein, under optimal conditions. This is now a recombinant protein. Small volumes of cell cultures will not yield a large amount of recombinant protein. Therefore, large-scale production is necessary to generate products that benefit humans. For this purpose, vessels called bioreactors are used.
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Answer:
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