人β-NGF基因在大肠杆菌中的表达  

The Study of Human β-NGF Gene Expressed in E.coli

作　　者：白艳艳[1] 杨吉成[1] 李丽娥[1] 盛伟华[1] 谢宇锋[1] 缪竞诚[1] 

机构地区：[1]苏州大学基础医学系细胞和分子生物学教研室,江苏苏州215007

出　　处：《苏州大学学报（医学版）》2004年第4期466-468,共3页Suzhou University Journal of Medical Science

基　　金：江苏省科技厅应用基础课题资助 (BJ990 70 )

摘　　要：目的　为 β NGF的基因制药选择一种新方法。 方法　直接从人血细胞基因组DNA中PCR扩增出 β NGF基因片段 ,将构建的重组体 pBV2 2 0 β NGF转化大肠杆菌DH5α,并将筛选的阳性克隆通过 4 2℃诱导表达。结果　经限制酶酶切电泳和DNA测序证实 ,确为 β NGF全长序列 (约 380bp) ;全菌经SDS PAGE电泳显示表达了 β NGF蛋白。结论　成功克隆了 β NGF基因全长序列并在大肠杆菌中获稳定表达。Objective To study a new method of expressing protein β-NGF through technique of gene recombination in vivo. Method A 380bp fragment was amplified from human blood genomic DNA by the polymerase chain reaction(PCR), and cloned into pUCm-T vector. The identified insert fragment from the recombinant pBV220-β-NGF was directedly ligated with linearized pBV220 with the compatible termini. E.coli DH5α was transformed with the expression recombinant pBV220-β-NGF and induced by heat. Results The cloned DNA fragment was identified as the full-length sequence encoding human β-NGF by restriction analysis and DNA sequencing. SDS-PAGE revealed the cloned NGF gene was expressed as a fusion protein in the cells transformed by pBV220-β-NGF. Conclusion The clone containing the full-length sequence encoding human β-NGF is obtained and successfully expressed in E.coli .

关 键 词：人基因组DNA 神经生长因子 大肠杆菌 基因表达 

分 类 号：R392.13[医药卫生—免疫学]