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.
出处
《微生物学通报》
CAS
CSCD
北大核心
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)
关键词
特异腐质霉
毕赤酵母
内切葡聚糖酶Ⅱ
分泌表达
Humicola insolens, Pichia pastoris, Endoglucanase Ⅱ, Secretive expression
