人血管抑素cDNA在酿酒酵母中的表达和分泌

Secretive Expression of Human Angiostatin cDNA in S. cerevisiae

目的 :在酿酒酵母中表达和分泌人血管抑素。方法 :应用DNA重组技术构建重组质粒 ;质粒DNA转化酵母用醋酸锂法 ;以纤溶酶原抗血清为第一抗体 ,采用ELISA方法检测表达产物。结果 :将MFα1信号肽和人血管抑素融合基因序列插入酵母表达载体pMA91的Bg1Ⅱ位点 ,构建了重组表达质粒 pMAA2 ,转化酿酒酵母GRF18后获得工程菌GRF18(pMAA2 ) ,其培养上清液ELISA分析阳性。结论 :人血管抑素在GRF18(pMAA2 )中得到表达 ,产物分泌至细胞外 ,能够与纤溶酶原抗血清发生特异的免疫反应。

Objective:To express and secrete human angiostatin in S. cerevisiae. Methods:DNA recombinant technique was used to construct recombinant plasmid,which was then transformed into yeast by lithium acetate method. The expression product was detected with ELISA. Results:The fused sequence of MFα 1 signal peptide coding region and human angiostatin cDNA was inserted into the Bg1 Ⅱ site on the shuttle vector pMA91 to form the plasmid pMAA2,which was then transformed into S. cerevisiae GRF18. ELISA analysis showed that the culture supernatant of the engineering train S. cerevisiae GRF18 (pMAA2) reacted positively with human plasminogen antisera. Conclusion:Angiostatin was successfully expressed in S. cerevisiae and secreted out to the culture.