猪丙型肝炎病毒核心蛋白结合蛋白6同源基因的克隆化研究

利用不同种属动物之间重要基因序列高度同源的理论，应用分子生物学与生物信息学技术和方法。克隆猪丙型肝炎病毒(HCY)核心蛋白结合蛋白6(HCBP6)的同源基因。首先应用酵母双杂交技术，以表达HCV核心蛋白的表达栽体作为曾饵，对于百万级的肝细胞cDNA文库酵母进行配合、筛选，首先获得人HCBP6的全长鳊码基因。然后应用美国国立生物工程中心(NCBI)建立的核苷酸序列数据库(CenBank)的同源基因的检索，搜索与之同源的来源于猪的表达序列标签(EST)。然后根据基因同泺性的原则，确定猪HCBP6的同源基因。获得了与HCC核心蛋白结合蛋白的36个基因片段，其中之一命名为HCBP6。根据基因同源性搜索，获得了来源于猪的EST基因序列片段。最终确立了猪HCBP6的同源基因。利用不同物种之间基因同泺性的原理、NCBI数据库GenBank同源基因的搜索，获得了猪HCBP6同源基因。生物信息学技术在后基因组时代具有重要地位和作用。

Cloning and sequencing analysis of Sus Scrofa hepatitis C virus core-binding protein 6(HCBP6) homologue gene

To clone Sus Scrofa hepatitis C virus (HCV) core protein_binding protein 6 (HCBP6) homologue gene by homologous gene search in the GenBank established by National Center of Bioengineering Institute (NCBI),National Institute of Hea1th (NIH),USA.Expression vector of HCV core protein was constructed.The transformed yeast was mated with yeast harboring hepatocyte cDNA library.Positive clones were selected by galactosidase activities and deficient medium growth screening.The homologous expressed sequence tag (EST) was found in the GenBank for Sus Scrofa cDNA fragments.The full length coding sequence for Sus Scrofa HCBP6 homologue was established by bioinformatics method.Thirty-six positive clones have been selected from yeast two_hybrid screening.Among them,there are 3 cDNA fragment without homology to any sequences in the GenBank.Huamn HCBP6 cDNA has been cloned. The Sus Scrofa HCBP6 full length coding sequence has been established by homologous search. Sus Scrofa HCBP6 full length coding sequence has been established by combination of molecular cloning and bioinformatics methods.Bioinformatics method is an important tool for identification and characterization of homologous genes from GenBank in the post_genomic era.