特异腐质霉中性内切葡聚糖酶Ⅱ基因的克隆及表达  

Cloning and Expression of the Endo-β-glucanaseⅡ Gene from Humicola insolens H31-3

作　　者：谭慧芳[1,2] 张国青[1] 郑光宇[3] 崔福绵[1] 钱世钧[1] 

机构地区：[1]中国科学院微生物研究所 [2]北京师范大学北京100875 [3]北京师范大学

出　　处：《微生物学通报》2006年第6期68-73,共6页Microbiology China

基　　金：国家十五科技攻关项目资助(No.2004BA713B03-01)

摘　　要：利用RT-PCR的方法,以特异腐质霉(Humicolainsolens)H31-3总RNA为模板,克隆到中性内切葡聚糖酶Ⅱ基因egl2的cDNA,将其插入到表达载体pGAPZαA中,重组质粒经线性化,电击转化毕赤酵母(Pichiapastoris)菌株GS115,筛选到分泌表达重组EGⅡ的毕赤酵母工程菌株。SDS-PAGE检测结果表明,重组EGⅡ在酵母中得到了特异性表达,表达产物的表观分子量约为55kD,同时对工程菌株的发酵条件和重组EGⅡ的性质进行了初步研究。A endo-β-glucanase Ⅱ （ EG Ⅱ） cDNA gene was isolated from the fungus Humicola insolens H31-3 by RT-PCR, It was cloned into the expression vector pGAPZαA, and the resultant recombinant plasmid was introduced into Pichia pastoris GS115by electroporation after lineared by BspH Ⅰ digestion, The recombinant Pichia pastoris strain was obtained and SDS-PAGE showed that the apparent molecular weight of the expression product was about 55kD. The culture condition and the properties of the recombinant EGⅡ were characterized elementarily.

关 键 词：特异腐质霉 毕赤酵母 内切葡聚糖酶Ⅱ 分泌表达 

分 类 号：Q78[生物学—分子生物学]