[Screening and cloning of the genes of protein interacting with the N-terminal protein of hepatitis B virus DNA polymerase by yeast-two hybrid technique]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005 Mar;19(1):84-6.
[Article in Chinese]


Objective: To screen and clone the genes of protein interacting with the N-terminal protein (TP) of hepatitis B virus DNA polymerase.

Methods: TP was amplified by polymerase chain reaction (PCR) and TP bait plasmid was constructed by using yeast two-hybrid system 3, then transformed into yeast AH 109. The transformed yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2 x YPDA medium. Diploid yeast was plated on synthetic dropout medium (SD/-Trp-Leu-His-Ade) and that containing X-alpha-GAL for selecting two times and screening. Plasmids were extracted from blue colonies, and sequence analysis was performed by bioinformatics.

Results: Forty-seven clones were obtained, these clones included human P36956 sterol regulatory element binding protein-1, RNA polymerase II subunit hsRPB7 mRNA, asialoglycoprotein receptor 2, transcript variant 3, ceruloplasmin (ferroxidase), transmembrane 4 superfamily member 2 and 19 of the hypothetical proteins and so on.

Conclusion: Genes encoding TP interacting proteins in hepatocytes were successfully cloned and the results suggest that TP has a wide variety of biological functions.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / metabolism*
  • Gene Library
  • Hepatitis B virus / enzymology*
  • Hepatitis B virus / genetics
  • Humans
  • Liver / metabolism
  • Plasmids / genetics
  • Protein Binding
  • Receptors, Virus / genetics
  • Receptors, Virus / isolation & purification
  • Receptors, Virus / metabolism
  • Transformation, Genetic
  • Two-Hybrid System Techniques*
  • Viral Proteins / chemistry
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*


  • Receptors, Virus
  • Viral Proteins
  • hepatitis B virus-binding protein, human
  • DNA-Directed DNA Polymerase