摘要
根据GenBank中发表的猪胃蛋白酶原A(pepsinogen)基因序列设计引物,采用RT-PCR技术,成功获得了猪胃蛋白酶原A基因,该基因全长1 240 bp,将该基因克隆到表达载体pPIC3.5K的EcoRⅠ和NotⅠ酶切位点构建重组表达质粒,通过电转化方法将重组质粒转化毕赤酵母GS115,检测结果表明猪胃蛋白酶原A基因在毕赤酵母中得到了表达.
The total RNA was extracted from swine gastric mucosa, which acted as the template to get a 1.24 kb segment of swine pepsinogen A gene cDNA by RT-PCR. The cloned cDNA was ligated into EcoR Ⅰ and Not Ⅰ sites of the vector pPIC3.5K downstream with the AOX1 alcohol oxidase promoter and transformed into Pichia pastoris GS115. The transformants grew in shaking flask and were induced by methanol, then analyzed by SDS PAGE, the results showed swine pepsinogen （EC 3.4.23.1） A gene was expressed successfully in Pichia pastoris GS115.
出处
《武汉大学学报（理学版）》
CAS
CSCD
北大核心
2006年第4期508-512,共5页
Journal of Wuhan University:Natural Science Edition
基金
湖北省科技厅重点攻关项目(99190503)
作者简介
乔宪凤（1968-），女，助理研究员，现从事生物工程研究．
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