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。
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 hepatocareinoma cell line HepG2, TDGF - 1 eDNA was amplified by RT - PCR using specific primers, then the eDNA was inserted into vector pHB to construct the recombinant vector pHB - TDGF - 1. The recombinant vector was transduced into escheriehia eoli 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. Conelusion： 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.
出处
《中国妇幼保健》
CAS
北大核心
2009年第35期5051-5053,共3页
Maternal and Child Health Care of China
基金
国家自然科学基金项目〔30670220〕
教育部科学技术研究重点项目〔107121〕
关键词
畸胎瘤衍化生长因子
TDGF-1
原核表达
Teratocarcinoma - derived growth factor - 1
TDGF - 1
Prokaryotic expression
