绿色木霉葡聚糖内切酶（EGⅠ）基因克隆及在酿酒酵母中的表达

摘要

In this paper,a 1400 bp gene from mycelium of industrial production strain Trichoderma viride Sn-9106 was cloned using RT-PCR with total RNA as templates.Compared with the sequence of E00390 endoglucanase from Genbank,the proportion of homologous amino acids was 91%.The EG I gene was constructed into expression vector of pESC and the recombinant EG I mature protein was expressed in Saccheromyces cerevisia strain Fm135a under the control of GAL10 promoter.After primary purification,specific activity of the recombinant endoglucanase was 2.45 U/mg.The SDS–PAGE analysis showed a band with apparent molecular weight of about 49 kD.%以纤维素酶工业生产菌株绿色木霉Sn-9106的总RNA为模板,采用RT-PCR技术获得大小为1 400 bp的cDNA片段,该序列与GenBank中的E00390葡聚糖内切酶序列相比,氨基酸序列同源性达91%。将EGⅠ基因构建入表达载体pESC中,在GAL10启动子控制下,目的基因在酿酒酵母Fm135a中成功表达,所得葡聚糖内切酶经初步纯化后,SDS-PAGE检测为49 kD,比活力为2.45 U/mg。