康氏木霉中调控纤维素酶形成的两个调控因子的克隆及功能性分析

Cloning and Functional Analysis of Two Regulators Involved in Regulation of Cellulase Formation in Trichoderma koningii

摘要:

锌指蛋白ACEI和Xyr1是里氏木霉中调控纤维素酶和木聚糖酶形成的两个调控因子, 它们能竞争性结合木聚糖酶基因xyn1的启动子。为进一步研究ACEI和Xyr1调控纤维素酶基因表达的机制, 本研究利用PCR技术扩增康氏木霉ACEI和Xyr1 DNA结合区的基因序列, 并使其在大肠杆菌中得到表达。凝胶迁移率移动试验表明ACEI和Xyr1的DNA结合区均可与纤维素酶基因cbh1启动子的287 bp序列特异性结合。提示ACEI与Xyr1不仅能竞争性结合xyn1启动子, 也能竞争性结合cbh1启动子。

Abstract:

ACEI and Xyr1 are two regulators that potentially involve in regulation of cellulases and xylanases formation in Trichoderma reesei, they compete for a binding site in the xyn1 (Xylanase1-encoding) gene promoter. To further investigate the mechanism for the transcriptional regulation of cellulases, DNA-binding domains of both ACEI and Xyr1 in T. Koningii were expressed from E. coli. They both showed bindings to the cbh1 promoter fragment (-304 bp to -18 bp) by electrophoresis mobility shift assays, suggesting ACEI and Xyr1 not only compete for binding to xyn1 promoter but also to cbh1 promoter.