摘要
为拓宽酵母菌的应用领域,构建具有分泌纤维素酶能力的酵母工程菌,利用质粒pPICZαA构建了康宁木霉纤维素酶cbh2基因的酵母表达载体,经线性化后采用电穿孔法转入毕赤氏酵母菌中.以甲醇为诱导物进行诱导,聚丙烯酰胺凝胶电泳检测结果证明该基因可成功表达.对重组蛋白进行了产酶活性测定,结果证明该重组蛋白具有纤维二糖水解活性,从而获得了一株可产生纤维素酶的酵母工程菌.
In order to widen the application field of yeast and to construct an engineering yeast strain that can produce cellulase, we constructed a yeast expression vector of cbh2 gene of Trichoderma koningii by use of pPICZαA. The vector was linearized and then transferred into Pichia pastoris by electroporation method. It was confirmed by SDS - PAGE that the gene was expressed successfully when being induced by methanol. We also detected the enzyme production activity of the recombinant protein in the transformed Pichia pastoris. The results show that the recombinant protein has the CBHII activity, thereby obtaining the engineering yeast strain capable of producing cellulase.
出处
《河南工业大学学报（自然科学版）》
CAS
北大核心
2010年第4期50-53,共4页
Journal of Henan University of Technology：Natural Science Edition
基金
河南工业大学校博士科研启动基金项目
河南省教育厅优秀青年骨干教师项目(155)
关键词
康宁木霉
酵母
cbh2基因
表达
Trichoderma koningii
Pichia pastoris
cbh2 gene
expression
