Cloning of pC/C and C gene of hepatitis B virus by T/A strategy

Ding Xie Liu, Kan Xian Luo

Research output: Contribution to journalArticlepeer-review

Abstract

A new strategy of cloning uncompatible DNA ends was reported here with the example of cloning of pC/C and C gene of hepatitis B virus. When the terminals of DNA fragments digested by restriction enzymes are blunt ends, they can be formed into tails with a single 3′ adenosine overhanged by modified with template-independent terminal transferase activity of Taq polymerase in reaction buffer containing dATP. Furthermore, if the DNA fragments have 5′-protruding sticky ends, these terminus can be filled into blunt ends with 5′ to 3′ Taq polymerase activity in the existence of dNTP, and then again added a single adenosine at their 3′ ends by the transferase activity of Taq polymerase. The fragments modified as such above-mentioned can be easily subcloned into T vector.

Original languageEnglish (US)
Pages (from-to)663
Number of pages1
JournalProgress in Biochemistry and Biophysics
Volume27
Issue number6
StatePublished - 2000
Externally publishedYes

Keywords

  • Gene recombinant
  • Hepatitis B virus
  • T vector
  • Taq polymerase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

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