【摘要】 目的:构建人畸胎瘤衍化生长因子(teratocarcinoma-derived growth factor-1,TDGF-1)原核表达载体,进一步探讨其生物学作用及机制。方法:从人肝癌细胞系HepG2中提取总RNA,利用TDGF-1特异性引物通过RT-PCR方法扩增TDGF-1 cDNA,克隆入pHB载体中,构建重组载体pHB-TDGF-1。将重组载体转化入大肠杆菌BL21(DE3)中进行表达,并通过SDS-PAGE及免疫印迹法对表达产物进行鉴定。结果:酶切及测序结果显示,重组质粒构建成功;SDS-PAGE及免疫印迹结果显示,所构建的重组载体可在大肠杆菌中高效表达TDGF-1。结论:该研究成功构建了人TDGF-1原核表达载体,并能够在大肠杆菌中有效表达TDGF-1,为进一步研究TDGF-1的生物学功能及其机制奠定了基础。更多还原

【Abstract】 Objective:To construct the prokaryotic vector of human teratocarcinoma-derived growth factor-1 (TDGF-1),explore the biological function and mechanism of TDGF-1.Methods:Total RNA was extracted from human hepatocarcinoma cell line HepG2,TDGF-1 cDNA was amplified by RT-PCR using specific primers,then the cDNA was inserted into vector pHB to construct the recombinant vector pHB-TDGF-1.The recombinant vector was transduced into escherichia coli BL21 (DE3) and the expression protein was detected by SDS-PAGE and Western blot.Results:Enzyme restriction digestion and sequencing results indicated that the recombinant vector was constructed exactly;SDS-PAGE and Western blot results showed that the recombinant prokaryotic expression vector could express TDGF-1 in escherichia coli efficiently.Conclusion:The prokaryotic expression vector pHB-TDGF-1 is constructed and expresses TDGF-1 in escherichia coli successfully,which establish the foundation for future research on TDGF-1 gene function and the mechanism.更多还原