作　　者： ; ; ; ;

机构地区： 广西大学生命科学与技术学院

出　　处： 《广西科学》 2007年第3期315-319,共5页

摘　　要： 用L-山梨糖诱导绿色木霉(Trichoderma viride)AS3.3711纤维素酶基因的转录,提取其总RNA反转录获得cDNA。PCR扩增葡聚糖内切酶(EG)和葡聚糖内切酶(EG)的cDNA基因,将其克隆到酵母载体中,构建产生纤维素酶的酿酒酵母工程菌。重组菌株能够识别EG和EG自身携带的信号肽而将表达产物分泌到胞外,故可采用刚果红平板染色法筛选具有羧甲基纤维素酶(CMCase)活性的重组转化子。重组菌株表达的EG酶活在诱导70h时达到最高(为0.08U/ml),表达的EG酶活在诱导60h时达到最高(为0.03U/ml)。 The cellulase genes from Trichoderma viride AS3. 3711 were induced with L-sorbose. Its total RNA was extracted. The cDNA was obtained by reverse transcription. The cDNA genes encoding endoglucanases Ⅰ and Ⅲ were isolated by PCR,and then they were cloned into a yeast vector to construct the cellulases-producing Saccharomyces cerevisiae engineering strains. Recombinant yeast strains can secret EGs Ⅰ and Ⅲ by the guide of the signal peptide themselves. So the expressions of EG I and EG Ⅲ can be screened by activity plate assays with Congo Red method respectively. The endoglucanase activity was assayed with CMC-Na as a substrate. The results showed that the enzyme activities of EG I and EG Ⅲ culminated at 0.08 U/ml and 0.03 U/ml respectively when they were induced for 70h and 60h respectively.

关 键 词： 绿色木霉 葡聚糖内切酶 酿酒酵母

领　　域： [生物学] [环境科学与工程]