title: Screening of hepatocyte proteins binding to F protein of hepatitis C virus by yeast two-hybrid system.
abstract: OBJECTIVE
  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 (alpha type) containing liver cDNA library plasmid in 2XYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-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-alpha-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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Pediatrics, the First Hospital of Xi'an Jiaotong University, Shaanxi Province, China. huangyp423@126.com
authors.forename: Yan-Ping
authors.initials: YP
authors.lastname: Huang
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.forename: Shu-Lin
authors.initials: SL
authors.lastname: Zhang
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.forename: Jiang
authors.initials: J
authors.lastname: Guo
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Yuan
authors.initials: Y
authors.lastname: Yang
authors.forename: Li-Ying
authors.initials: LY
authors.lastname: Zhang
authors.forename: Gui-Qin
authors.initials: GQ
authors.lastname: Bai
authors.forename: Xue-Song
authors.initials: XS
authors.lastname: Gao
authors.forename: Dong
authors.initials: D
authors.lastname: Ji
authors.forename: Shu-Mei
authors.initials: SM
authors.lastname: Lin
authors.forename: Qing
authors.initials: Q
authors.lastname: Shao
mesh_terms: D045744:Cell Line, Tumor; D015723:Gene Library; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D014758:Viral Core Proteins / Q000378:metabolism*
chemical_list: D014758:Viral Core Proteins; C433031:hepatitis C protein F, Hepatitis C virus