摘要
构建组织因子表达质粒并转染小鼠骨髓瘤细胞SP2/0,分泌表达有活性的组织因子。用阳离子聚合物法将构建好的转染质粒pIRESneo3-TF1-218转染入SP2/0细胞,经G418抗性筛选后,蛋白可由SP2/0细胞无血清培养分泌表达。G418 800ug/ml筛选到的表达克隆,经无血清培养,可分泌表达,western blot证明其为人组织因子衍生物,Mr为40k,脂化后,具有促凝血活性。成功构建组织因子蛋白表达质粒pIRESneo3-TF1-218。
Soluble tissue factor （sTF） was successfully expressed and secreted in an active form by transfecting the expression plasmid construct containing sTF gene into mouse SP2/0 cells. Plasmid pIRESneo3-TF1218 was constructed and transfected into SP2/0 cells and selected by G418. When the SP2/0 cells grow in serum free medium sTF is expressed and secreted into the medium. The clones growing under 800ug/ml G418 were cultured in serum free medium. The expressed sTF was secreted into the medium and identified by Western Blot with MW 40kDa. After being relipidated the sTF contain medium demonstrated coagulation activity.
出处
《药物生物技术》
CAS
CSCD
2009年第3期194-197,共4页
Pharmaceutical Biotechnology
