生物技术通报Issue(11)：140-144,5.
地鳖虫纤溶活性蛋白cDNA序列克隆与毕赤酵母表达
Cloning and Expression of Eupolyphaga sinensis Walker Fibrinolytic Protein cDNA in Pichia pastoris
摘要
Abstract
The primers were designed corresponding to Eupolyphaga sinensis Walker fibrinolytic protein (EFP) cDNA sequence which has been reported. The cDNA sequence encoding fibrinolytic enzyme was cloned from fresh body of Eupolyphaga sinensis Walker by RT-PCR,and then was blasted against other fibrinolytic enzyme cDNA sequences in the Genbank. The analysis of the sequence data indicated that the codeing region of cDNA fragment, which encoded 224 amino acid residues,was about 675 bp in size. The amplified cDNA fragment was cloned into the eukaryotic expression vector pPICZα-A, to produce the expression vector pPICZα-A-EFP. The re-combinant plasmid was trabsformed into P. pastoris. EFP fusion protein was obtained after the addition of methanol into the growth media. SDS-PAGE analysis revealed that the EFP was expressed . A protein band of 28. 2 kD appeared on SDS-PAGE gel. rEFP was secreted into the culture medium and was able to dissolve artificial fibrin plates.关键词
地鳖虫/纤溶活性蛋白基因/cDNA克隆/毕赤酵母表达Key words
Eupolyphaga sinensis Walker/Fbrinolytic protein/cDNA clone/Ekaryotic expression分类
生物科学引用本文复制引用
李兴暖,韩雅莉..地鳖虫纤溶活性蛋白cDNA序列克隆与毕赤酵母表达[J].生物技术通报,2009,(11):140-144,5.基金项目
国家自然科学基金 (30772739),博士后科学基金(20080430850) （30772739）
