瑞氏木霉内切葡聚糖酶基因在酿酒酵母中的表达研究

将PCR合成的瑞氏木霉 (Trichodermareesei) β 内切葡聚糖酶I(EGI)cDNA基因片断分别插入酵母met1 0和 pgk1启动子和终止子序列之间 ,构建了在不同启动子控制下 ,具有不同拷贝数的eg1表达分泌质粒pRS41 5ME、pRS41 5PE和 pRS42 5PE。通过电转化使重组质粒转移至实验室酿酒酵母H1 5 8菌株中 ,分别得到了 3株酵母转化子H1 p、H2p和H1m。在 3株酵母转化子中 ,重组 β 内切葡聚糖酶I都能在酶自身信号肽序列引导下进行分泌型表达。在YPD培养基中 3株重组酵母生长速率大致相同 ,H1m ,H1 p与H2 p的内切葡聚糖酶活力分别为 70 .4,1 2 6.7和 1 2 5 .0U/mL。

Expression of EGI from Trichoderma Reesei in Saccharomyces cerevisiae

To study the effects of different promoters and copy numbers, the eg1 gene from the filamentous fungus Trichoderma reesei was inserted between the promoter and terminator regions of either the pgk1 or met10 gene of yeast. The eg1 gene was transferred to Saccharomyces cerevisiae on a multicopy plasmid or a CEN/ARS plasmid, and was expressed in recombinant S.cerevisiae H1m, H1p and H2p.Three recombinant S.cerevisiae yeast strains with different β-endoglucanase activities were constructed.