可溶性重组人组织因子(rhTF_(243))分泌型表达载体的构建和表达  

Construction and Expression of Excreted Express Carrier of Recombinant Soluble Human Tissue

作　　者：赵邑[1] 齐延红[1] 潘薇薇[1] 卢晓霞[1] 刘彦萍[1] 赵峰梅[1] 

机构地区：[1]山西省生物研究所,山西太原030006

出　　处：《生物技术》2007年第3期15-17,共3页Biotechnology

基　　金：山西省自然科学基金项目资助("人重组组织因子基因工程菌的构建与表达";编号:20051083)

摘　　要：目的:构建含有TF的胞外区和跨膜区基因的phoA-TF243分泌型表达载体,在大肠杆菌中以可溶形式表达重组人组织因子(rhTF243)。方法:从人胎盘组织中提取总RNA,以RT-PCR扩增获得目的基因,并克隆到phoA载体中,在大肠杆菌MM294中表达rhTF243,产物采用免疫亲合层析进行纯化。结果:通过低磷酸盐诱导工程菌,获得重组人组织因子(rhTF243) ,表达水平占菌体总蛋白量的16 .3 %,经免疫亲合层析纯化,产物纯度达到95 %以上,分子量为27 .4kD。结论:获得了rhTF243,具有与完整分子完全相同的凝血功能。Objective： To construct excreted prokatyotie expression vector and express the recombinant human TF243 （rhTF243） which contains only the extracelluar domain and transmembrane domain of TF in E. coli. Methods： The eDNA gene of human placental total RNA and cloned into phoA vector, then expressed with E. coli. MM294.Results： After induced by low concentration phoephate, rhTF243 can be obtained and accounted for about 16.3% of the total cellular protein. Further more the recombinant protein is soluble and can be purifieated by immunity affinity chromatography pillar. The content of the purifacated protein can be over 95%. Conclusion： Human TF243 was obtained successfully. The recombinant soluble tissue have better coagulant activity existing of phospholipids in prothrombine time test.

关 键 词：重组人组织因子 基因克隆 大肠杆菌 分泌表达 

分 类 号：Q786[生物学—分子生物学]