Question

Question.If a desired gene is identified in organism for some experiments, explain the process of the following: 1) Cutting this desired gene at specific location. 2)Synthesis of multiple copies of this desired gene.

Answer 1

Hi there,

1.Cutting of DNA at specific locations is done by using restriction enzymes. The purified DNA is incubated with the specific restriction enzyme at conditions optimum for the enzyme to act..

2.(i) Isolation of the genetic material (DNA) is carried out in the following steps:

(a) DNA is enclosed within the membranes. To release DNA along with other macromolecules such as RNA, proteins, polysaccharides and lipids, bacterial cells/plant or animal tissue are treated with enzymes such as lysozyme (bacteria), cellulase (plant cells), chitinase (fungus)..

Hope it has Help!

Answer 2

(i)

• Identifying the restriction endonuclease that recognises the palindromic nucleotide sequence of the desired gene.
• The restriction endonuclease inspects the DNA sequences
• Finds and recognises the site
• Cuts each of the double helix at the specific point
• A little away from the centre of the palindromic site
• Between the same two bases on the opposite strand.
• Makes the over hanging stretch single stranded portion as a sticky end.

(ii)

• By PCR/Polymerase Chain Reaction
• Desired gene is synthesised in vitro
• DNA is denatured
• Annealed using two sets of primers
• Thermostable Taq polymerase extends the primers using nucleotides (provided in the reaction and genomic DNA as template)
• Amplified fragments are ligated.