【作者】 李娜； 赵明文；

【Author】 Li Na Zhao Ming-Wen (Key Laboratory of Microbiological Engineering of Agricultural Environment of Ministry of Agriculture; College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, P. R.China)

【机构】 南京农业大学生命科学学院农业部农业微生物环境工程重点开放实验室；

【摘要】 以酿酒酵母鲨烯单加氧酶基因的全长质粒DNA为模板进行PCR扩增;将PCR产物克隆到巴斯德毕赤酵母Pichia pastoris表达载体pPIC9K中,并位于α-因子信号肽序列的下游,获得重组质粒pPIC9K-SE。重组质粒线性化后用电转化法导入毕赤酵母菌株GS115中,经大量筛选,及PCR验证获得在染色体上整合有目的基因的菌株,取上清液进行SDS-PAGE 电泳可以检测到目的条带。更多还原

【Abstract】 The expression of squalene epoxidse in Pichia pastoris was investigated. The DNA encoding SE was cloning by PCR and the PCR product was inserted into the vector pPIC9K, downstream of a -factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-SE was lineared by Bg1II digestion and introduced into the host P. pastoris GS115 by electroporation. After screen, the recombinant P. pastoris stains were obtained. The results of SDS-PAGE proved that the recombinant plasmid pPIC9K-SE was secreted into culture medium.更多还原