里氏木霉木聚糖酶基因XYN2的克隆与表达  

Cloning and expression of the T. reesei xylanase gene XYN2

作　　者：王向明[1] 欧阳嘉[2] 严明[1] 许琳[1] 

机构地区：[1]南京工业大学制药与生命科学学院,江苏南京210009 [2]南京林业大学化学工程学院,江苏南京210037

出　　处：《南京工业大学学报（自然科学版）》2007年第5期82-87,共6页Journal of Nanjing Tech University(Natural Science Edition)

基　　金：国家重点基础研究发展计划("973"计划)资助项目(2003CB615701);国家自然科学基金资助项目(20336010)

摘　　要：采用RT-PCR(reverse-transcription PCR assay)技术,从里氏木霉QM9414 mRNA中扩增出木聚糖酶基因XYN2,并与质粒pYX212相连,构建了组成型表达载体pYX212/XYN2.将此重组质粒转化入酿酒酵母菌株YPH499进行表达.分析了不同碳源、温度、摇瓶转速、初始pH对其产酶的影响.实验结果表明,最佳碳源是葡萄糖,最佳转速是260 r/min,最佳初始pH是6.0.优化重组菌的发酵条件后得到的最大酶活为0.55 IU/mL.XYN2 gene fragments were successfully amplified from Trichoderma reesei strain QM9414 mRNA by RTPCR （reverse-transcription PCR assay） technology and were ligated to pYX212 vector as the expression vector pYX212/XYN2. Saccharornyces cerevisia YPH499 competent cells were transformed with the recombinant plasmid and began expressing. The effects of different carbon sources, temperature, rotary speed and initial pH on xylanase production of the recombinant were invrstigated. The result indicats that the optimal carbon source is glucose, the optimal temperature is 30 ℃, the optimal rotary speed is 260 r/min, and the optimal initial pH is 6.0. The maximum xylanase activity of the recombinant is 0.55 IU/mL after optimizing the fermentation condition.

关 键 词：RT-PCR TRICHODERMA REESEI XYN2基因 酿酒酵母 克隆 表达 

分 类 号：Q786[生物学—分子生物学]