Q.Restriction endonuclease was isolated for the first time by W. Arber in 1962, in bacteria. DNA, the genetic material can be manipulated by addition or substitution of the desired gene by using a specific restriction endonuclease resulting in rDNA. This is one of the major steps in biotechnology.
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Start your 14-day free trial to unlock the full solution →First isolated restriction enzyme: Hind II. 'Restriction' because it restricts phage growth by cutting foreign DNA. It works by recognising a specific palindromic sequence and cutting the DNA backbone of both strands at fixed points inside it.
i) First isolated restriction endonuclease: Hind II. (Although the restriction phenomenon was studied by W. Arber, the first restriction enzyme to be actually isolated and characterised was Hind II.)
ii) Why called 'restriction' endonucleases: In bacteria these enzymes restrict the growth (multiplication) of invading bacteriophages and other foreign DNA by cutting the foreign DNA into pieces, thus acting as part of the bacterial defence system. Because they act within the DNA molecule (not from the ends), they are endonucleases.
iii) How a restriction endonuclease functions:
- It inspects the length of a DNA and recognises a specific base sequence — usually a short palindromic sequence (reads the same 5′→3′ on both strands).
- It binds to this recognition site.
- It then cuts the sugar-phosphate backbone of both strands at specific points within (or near) that sequence — often making a staggered cut that leaves single-stranded sticky ends (or a straight cut giving blunt ends).
- The bacterium's own DNA is protected from being cut by methylation of the recognition sites. …
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