【摘要】 巴斯德毕赤酵母是一种很有潜力的真核表达体系。本文报告将已克隆的乙型肝炎病毒ｐｒｅＳ２－Ｓ基因亚克隆于巴斯德毕赤酵母胞内表达质粒ｐＰＩＣ３，构建重组ｐＰＩＣ３／ｐｒｅＳ２－Ｓ质粒，经酶切线性化后，将其用电转化导入酵母细胞内，经ＰＣＲ及ｄｏｔｂｌｏｔ检测，挑选出在染色体上稳定整合了乙肝病毒包膜中蛋白编码基因的嗜甲醇毕赤酵母菌株，为高效表达乙肝病毒包膜中蛋白奠定了基础更多还原

【Abstract】 The recombinant plasmid pPIC3/preS2 S was amplified by transforming E.coli TOP10F’ after subcloning the preS2 S gene of Hepatitis B Virus into Pichia pastoris intracellular expression vector pPIC3.Purified pPIC3/preS2 S were linearized with restriction enzyme SacⅠ and Bgl Ⅱ respectively and then were introduced into Pichia pastoris GS115 strain by means of electroporation.Stable transformants of Pichia pastoris were generated via homologous recombination between the vector and regions of homolgy within the genome.Ten HIS +MUT + and twelve HIS +MUT S Pichia transformants were conformed the integration of preS2 S gene by PCR and dot blot analysis.更多还原