人源性肝细胞生长刺激因子cDNA克隆和序列分析

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础。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 6 0 0bp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 74 8bp ,含有 5 94bp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 1 98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础

Cloning and sequence of human hepatocyte DNA synthetic stimulated factor cDNA

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 degenerating PCR, labelled the fragment as a probe to screen human fetal liver cDNA library, and then analyzed the HDSSF gene sequence. Results A 600 base pair fragment was obtained and labelled successfully with DIG. Human HDSSF sequence was determined by screening the library and gene sequencing. Its cDNA chain was composed of 748 base pairs including 594 base pair open readcode frame and 5′ and 3′ terminal noncode district sequence. Conclusion By screening cDNA library, the HDSSF gene cloning has been gained successfully. HDSSF should be a new hepatocyte growth substance which consists of 198 amino acid residues.