人源性肝细胞生长刺激因子cDNA 克隆和序列分析 - 中南大学学报.PDF

BULL HUNAN MED UNIV 2003, 28( 3) 209 cDNA 1 2 唐世刚 , 苏先狮 ( 1. 中南大学湘雅三医院感染病科, 长沙410013; 2. 中南大学湘雅二医院传染科, 长沙410011) [] 目的: 阐明人源性肝细胞生长因子(HDSSF) 的本质,为进一步基因克隆奠定基 方法: 以简并PCR 扩增获取目的基因片断;杂交筛选人胎肝 cDNA 文库; 末段终止法进行HDSSF 核苷酸序列分析结果:简并 PCR 扩 增出接近HDSSF 分子量 600 bp 片断; 人胎肝 cDNA 文库经初筛和复筛获得含有目的基因的单克隆噬斑; 测定了 HDSSF 基因序列,其 cDNA 链为748 bp,含有594 bp 的完整开放读码框架和5 及3 末端非编码区序列序列分析显 示HDSSF 应为一含有 198 个氨基酸残基的新的促肝细胞生长物质结论: 本研究成功获得目的基因HDSSF 克隆, 人HDSSF 克隆成功为开展人源性HDSSF 的重组产品研制奠定了基 [] 肝细胞生长因子; 人; 克隆; 序列分析 [] R392.4 [ ] A [ ] 2003) Cloning and sequence of human hepatocyte DNA synthetic stimulated factor cDNA 1 2 TAN Shigang , SU Xianshi ( 1. Dep artment of Inf ectious Diseases, Third Xiangya H ospital, Central South University , Changsha 410013; 2. Dep artment of Inf ectious Diseases , Second Xiangy a H osp ital, Central South University, Changsha 410011, China Abstract: Objective To clarify the nature of human hepatocyte DNA synthetic stimulated factor (HDSSF) and to lay a foundation for its gene cloning and expression. Methods We amplified the target gene fragment by degeneratingPCR, labelled the fragment as aprobeto screen human fetal liver cDNA library, and then analyzed the HDSSF gene sequence. Results A 600 base pair fragment was obtained and labelled suc cessfullywith DI . Human HDSSF sequencewas determined by screeningthe library and gene sequencing. Its cDNA chain was composed of 748 base pairs including 594 basepair open readcodeframe and 5 and 3 termi nal noncode district sequence. Conclusion By screen