中山大学学报（自然科学版）2012，Vol.51Issue(2)：86-90,96,6.
花鳗鲡重组促黄体生成素在毕赤酵母中的表达
Expression of A Recombinant Luteinizing Hormone of Marble Eels,Anguilla marmorata Using Methylotropic Yeast, Pichia pastoris
摘要
Abstract
The GTHα、LHβ gene fragments were amplified by PCR, which the template was pituitary cDNA of marble eels. In the way of bridging, the two fragments were connected to a single gene LHβα, a linker inserted between them. The LHβα gene fragment was subcloned to pPICZαA with double enzyme digestion and connection, constructing eukaryotic co-expression plasmid pPICZαA-LHβα. The recombi-nant plasmid was transfected into a native yeast strain X-33 by electrotransformation. The high-copy clones were selected by the YPDS plates containing different concentrations of Zeocin. The positive clones were induced by 0.7% methanol and expression products were tested by SDS-PAGE and western blot, and its molecular weight was determined as about 45 000.关键词
花鳗鲡/促黄体生成激素/linker/共表达载体pPICZαA-LHβα/毕赤酵母Key words
Anguilla marmorata/ luteinizing hormone/ linker/ co-expression plasmid pPICZαA-LHβα/ Pichia pastoris分类
生物学引用本文复制引用
毛黎敏,张勇,李阳源,黄海,李波,李文笙,陈国华,林浩然..花鳗鲡重组促黄体生成素在毕赤酵母中的表达[J].中山大学学报（自然科学版）,2012,51(2):86-90,96,6.基金项目
国家自然基金面上资助项目(30770283) （30770283）
广东省教育部产学研结合资助项目(2010B090400551):江苏省科技成果转化专项资金资助项目(BA2009083) （2010B090400551）
海南省教育厅高等学校科学研究资助项目(Hjkj20082 22) （Hjkj20082 22）
