里氏木霉纤维二糖酶bglII基因的cDNA克隆及其在大肠杆菌中的表达  

CDNA CLONING CELLOBIASE BGLII GENE FROM TRICHODERMA REESEI AND ITS EXPRESSION IN ESCHERICHIA COLI

作　　者：陈新爱[1] 夏黎明[1] 岑沛霖[1] 

机构地区：[1]浙江大学生物工程研究所,杭州310027

出　　处：《菌物系统》2002年第2期223-227,共5页Mycosystema

基　　金：浙江省科委国际合作项目

摘　　要：将里氏木霉总RNA反转录得到cDNA第一链,并以之为模板进行 RT-PCR合成约1.4kb的纤维二糖酶基因cDNA,将所得的cDNA经测序后克隆到温度敏感型表达载体pJW2中,经PCR和双酶切鉴定筛出阳性重组子,温度诱导后,经酶活测定,bgl II基因在大肠杆菌中得到了表达,酶活为0.75IU/mL。Cellobiase is a hydrolytic enzyme which acts on various compounds with b-D-glucosidic linkages. It plays a key role in hydrolyzing cellobiose to glucose. Cellobiase bglII gene was cloned by RT-PCR using first strand cDNA as template after reverse transcripting T.reesei total RNA. After sequenced, the bglII cDNA was then inserted directionally into a temperature-inducible prokaryotic expression vector pJW2. The bglII gene was expressed in E.coli DH5a.

关 键 词：里氏木霉 纤维二糖酶 bg1Ⅱ基因 CDNA克隆 大肠杆菌 表达 原核表达 

分 类 号：Q78[生物学—分子生物学]