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

作　　者：凌敏[1,2] 覃拥灵[1] 李楠[1] 梁智群[1] 

机构地区：[1]广西大学生命科学与技术学院亚热带生物资源保护和利用重点实验室,南宁530004 [2]广西医科大学生物化学与分子生物学教研室,南宁530021

出　　处：《生物工程学报》2008年第10期1808-1812,共5页Chinese Journal of Biotechnology

基　　金：国家自然科学基金资助项目(No.20066001);广西亚热带生物资源保护和利用重点实验室开放课题(No.SB0709)资助~~

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。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 （Xylanasel-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 cbhl 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 cbhl promoter.

关 键 词：调控因子 纤维素酶 凝胶迁移率移动试验 

分 类 号：Q78[生物学—分子生物学]