title: Screening of genes of proteins interacting with p7 protein of hepatitis C virus from human liver cDNA library by yeast two-hybrid system.
abstract: OBJECTIVE
  To investigate the biological function of p7 protein and to look for proteins interacting with p7 protein in hepatocytes.
  
  
  METHODS
  We constructed p7 protein bait plasmid by cloning the gene of p7 protein into pGBKT7, then transformed it into yeast AH109 (a type). The transformed yeast was mated with yeast Y187 (alpha type) containing liver cDNA library plasmid, pACT2 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 of plasmids from blue colonies, we performed sequence analysis by bioinformatics.
  
  
  RESULTS
  Fifty colonies were selected and sequenced. Among them, one colony was Homo sapiens signal sequence receptor, seven colonies were Homo sapiens H19, seven colonies were immunoglobulin superfamily containing leucine-rich repeat, three colonies were spermatid peri-nuclear RNA binding proteins, two colonies were membrane-spanning 4-domains, 24 colonies were cancer-associated antigens, four colonies were nucleoporin 214 ku and two colonies were CLL-associated antigens.
  
  
  CONCLUSIONS
  The successful cloning of gene of protein interacting with p7 protein paves a way for the study of the physiological function of p7 protein and its associated protein.
authors.affiliation: Department of Pediatrics, The First Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi Province, China. huangyp423@126.com
authors.forename: Yan-Ping
authors.initials: YP
authors.lastname: Huang
authors.forename: Shu-Lin
authors.initials: SL
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
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: Yan-Wei
authors.initials: YW
authors.lastname: Zhong
authors.forename: Qing
authors.initials: Q
authors.lastname: Shao
mesh_terms: D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D015723:Gene Library; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism* / Q000821:virology*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D000086082:Viroporin Proteins
chemical_list: D004279:DNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; C088369:p7 protein, Hepatitis C virus; D000086082:Viroporin Proteins