摘要
为探讨快速高效表达山羊FSH(促卵泡素)基因的方法,将获得的FSHα,β亚基基因克隆到毕赤酵母快速表达载体pPICZαA中,然后将构建的载体线性化处理后,电击转化至毕赤酵母GS115中,经Zeocin抗生素筛选后获得阳性菌落,经甲醇诱导表达后进行SDS-PAGE和Western–blot分析。结果表明:pPICZαA载体可以快速正确的表达FSH,分子量为27KD,放射免疫法测定表达上清,表达量为20.155 mIU/mL,显著高于本实验室在细胞中的表达。
In order to discuss., the methods of expression of goat follicle-stimulating hormone fastly and efficiently.The obtained FSH α, β-subunit genes were cloned into Pichia pastoris pPICZ α A vector,then the recombined vector pPICZ α A FSH or, β were lined and transformed into Pichia pastoris GSl15 by electroporation.The positive Yeast transformants were screened by the medium containedZeocin.After methanol induction,the supernatant was analysised by SDS-Polyacrylamide Gen Electrophoresis and Western-blot.The results show that FSH could be expressed fastly and precisely and the molecular weight is about 27KD.The concentration of supernatant was detected by Radio-immunoassay and the average expression is 20.155mlU/mL ,which is higher than expressed in cells significantly.
出处
《黑龙江动物繁殖》
2011年第5期12-15,共4页
Heilongjiang journal of animal reproduction
关键词
山羊
FSH基因
毕赤酵母表达载体
表达
goat follicular-stimulating hormone gene
Pichia pastoris vector
expression
