摘要
将大口黑鲈(Micropterus salmoides)抗菌肽hepcidin cDNA定向插入真核表达载体pPICZαA,通过SacI酶切线性化重组表达质粒pPICZαA-hepcidin,电转化毕赤酵母P.pastoris GS115,PCR扩增筛选,甲醇诱导表达等进行了hepcidin真核表达工程菌株的构建及诱导表达。结果显示:hepcidin cDNA成功插入表达载体pPICZαA,并整合到酵母基因组中;经甲醇诱导,RT-PCR检测显示hepcidin cDNA在酵母细胞中成功转录。
Ab In this research, largemouth bass （Micropterus salmoides） hepcidin cDNA was cloned into the eukaryotic expression veαor pPICZαA. The linearized recombinant plasmid pPICZαA-hepcidin was transformed into yeast P. pastoris GS115 by eleαroporation. Hepcidin gene eukaryotie expression engineering yeast was screened out by PCR deteαion and expressed by methanol induαion. These results showed that largemouth bass hepcidin gene was inserted into expression veαor pPICZαA, and then integrated into yeast genome. After the induαion expression, reverse transcription （RT）-PCR deteαion revealed that hepcidin was successfully transcribed by recombinant yeast host strain GS115.
出处
《淡水渔业》
CSCD
北大核心
2009年第2期71-75,共5页
Freshwater Fisheries
基金
国家科技基础条件平台工作项目(2005DKA21103)
广东省重大科技兴渔项目(B200701A06)
