内毒素结合肽表达质粒的构建及其在大肠杆菌中的表达  

RECOMBINATION AND EXPRESSION OF ENDOTOXIN BINDING PEPTIDE IN E.COLI

作　　者：麻莉[1] 刘友生[1] 王晓东[1] 葛晓冬[1] 李永旺[2] 

机构地区：[1]第三军医大学西南医院 [2]第三军医大学新桥医院

出　　处：《解放军医学杂志》2003年第4期302-304,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金资助课题 (编号 30 0 0 0 1 71 )

摘　　要：应用PCR技术 ,从含人NH·LBPcDNA的质粒中扩增出 93bp的内毒素结合肽 (endotoxinbindingpeptide,EBP)基因片段 ,插入原核融合蛋白表达质粒PinpointXa3的多克隆位点构建成表达质粒 ,转化大肠杆菌DH5α,筛选得到阳性重组子PinpointXa3 EBP并进行酶切鉴定及序列分析。异丙基硫代半乳糖苷 (IPTG)诱导重组融合蛋白的表达 ,经SDS PAGE及Westenblot鉴定得到分子量约为 16 .5kD的生物素化EBP融合蛋白 ,为进一步研究其内毒素拮抗效应打下良好基础 。Ninty three bp endotoxin binding peptide gene was cloned with PCR from the amino terminal of human LBP cDNA, which was introduced into the expression plasmid Pinpoint Xa 3. After the constructs being transformed into E.coli DH5α, the postive recombinants were identified by restriction enzyme analysis and DNA sequencing. Induced by IPTG, the desired 16.5kD EBP fusion protein was expressed and confirmed with SDS PAGE and Western blotting. These results paved a reasonable way for the study of a new endotoxin binding peptide.

关 键 词：内毒素结合肽 表达 质粒 构建 大肠杆菌 融合蛋白 

分 类 号：R378.21[医药卫生—病原生物学]