猪PDCD5基因的克隆与真核表达

用RT-PCR方法从猪肝组织中扩增出猪PDCD5(programmed cell death 5)编码序列,TA克隆至pMD-19T载体中,软件分析猪PDCD5基因的核算序列和蛋白序列,进行染色体定位.构建真核表达载体,将PDCD5基因编码区序列插入绿色荧光蛋白报告基因的真核表达载体pEGFP-C1中.通过脂质体转染法将重组载体瞬转入猪脐静脉血管内皮细胞系(SUVECs)进行瞬时表达.结果表明:该序列编码125个氨基酸,猪PDCD5基因定位于猪6号染色体,含有6个外显子,与人PDCD5基因高度同源.双酶切鉴定和测序表明:重组真核表达载体构建成功,荧光检测和Western blot检测显示PDCD5融合蛋白表达.研究结果为探讨猪PDCD5基因在细胞凋亡调控中的功能提供了基础数据.

Molecular Cloning and Eukaryotic Expression of Porcine PDCD5 Gene

An experiment was made to provide basic data for the study of the functions of PDCD5 gene in apoptosis regulation.Porcine PDCD5 gene was obtained by RT-PCR from porcine liver tissue,and the PCR product was cloned into T-easy vector pMD-19T.A software was used to analyze its nucleic acids and protein sequences,and the chromosomal location of porcine PDCD5 was determined.Then the PDCD5 gene was subcloned into the eukaryotic expression vector pEGFP-C1 of the green fluorescent protein(GFP) reporter gene encodin...