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For insertion of DNA into a suitable vector several methods are used and homopolymer tailing is one of them. Which specific enzyme is used in this method to synthesize homopolymer tail ?

  • A.\(T_{4}\) DNA ligase
  • B.Polynucleotide kinase
  • C.Terminal deoxynucleotidyl transferase
  • D.\(S_{1}\) nuclease

Answer: C

Homopolymer tails are made by Terminal deoxynucleotidyl transferase.

Terminal transferase adds nucleotides to the 3'-OH ends of DNA without a template. Given only one deoxynucleotide, for example dCTP, it builds a tail of that base alone. In homopolymer tailing the vector is tailed with one base (say poly-dG) and the insert with the complementary base (poly-dC), so the two anneal through their tails.

T4 DNA ligase joins DNA ends by forming phosphodiester bonds but does not add nucleotides. Polynucleotide kinase adds a phosphate to 5'-OH ends. S1 nuclease removes single-stranded DNA, which would trim tails rather than build them.

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