Screening of hepatocyte proteins binding to F protein of hepatitis C virus by yeast two-hybrid system

Authors:
  • JCJun Cheng
    Jun Cheng
  • SZShu-Lin Zhang
    Shu-Lin Zhang
  • LWLin Wang
    Lin Wang
  • JGJiang Guo
    Jiang Guo
  • YLYan Liu
    Yan Liu
  • YYYuan Yang
    Yuan Yang
  • LZLi-Ying Zhang
    Li-Ying Zhang
  • GBGui-Qin Bai
    Gui-Qin Bai
  • XGXue-Song Gao
    Xue-Song Gao
  • DJDong Ji
    Dong Ji
  • SLShu-Mei Lin
    Shu-Mei Lin
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Abstract
AIM: To investigate the biological function of F protein by yeast two-hybrid system. METHODS: We constructed F protein bait plasmid by cloning the gene of F protein into pGBKT7, then recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-α-gal for selection and screening. After extracting and sequencing plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics. RESULTS: Thirty-six colonies were selected and sequenced. Among them, 11 colonies were zymogen granule protein, 5 colonies were zinc finger protein, 4 colonies were zinc-α-2-glycoprotein, 1 colony was sialyltransferase, 1 colony was complement control protein factor I, 1 colony was vitronectin, and 2 colonies were new genes with unknown function. CONCLUSION: The yeast two-hybrid system is an effective method for identifying hepatocyte proteins interacting with F protein of hepatitis C virus. F protein may bind to different proteins.

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