摘要
【目的】构建水貂生长激素(mGH)基因真核表达载体,并在巴斯德毕赤酵母GS115中进行表达,为大量获得水貂生长激素奠定基础。【方法】将体外合成的水貂生长激素基因插入分泌型表达载体pPIC9K。将重组的pPIC9K-mGH用SacI酶切后,LiC1法转化巴斯德毕赤酵母菌株GS115,经G418筛选后进行甲醇诱导表达,并以SDS-PAGE和Western blot检测酵母上清中水貂生长激素的表达。【结果】酵母上清中可见与目的蛋白相对分子量(31kd)相符的条带,该条带可被水貂生长激素多克隆抗体特异识别。【结论】正确构建了水貂生长激素真核表达载体,该蛋白能在巴斯德毕赤酵母中成功表达。
[Objective]To construct eukaryotic expression vector of Mustela vision growth hormone（mGH）, and express in Pichia Pastoris strain GS115, make foundation to gain amounts of Mustela vision growth hormone. [Method]Mustela vision growth hormone, which was chemically synthesized, was inserted into secretion vector pPIC9K. The recombined pPIC9K - mGH was digested by Sac I and transferred into Pichia Pastoris strain GS115 with LiCI. After screening with G418, the positive yeast clone was induced to express with methanol, and the expression product was analyzed with SDS-PAGE and Western blot. [Results]A positive protein band with molecular weight about 31 kD was found in the expression product, which is accordant with interest protein, and this band could be specifically recognized by anti-mGH polyclonal antibody. [Conclusion]The gene of mGH was cloned into the expression vector correctly and was expressed successfully in P. pastoris expression system.
出处
《中国农学通报》
CSCD
2008年第3期30-34,共5页
Chinese Agricultural Science Bulletin
基金
山东信得药业企业自主课题
