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第 29卷第 5期南京工业大学学报 Vol. 29 No. 5
2007年 9月 JOURNAL OF NANJ ING UN IVERSITY OF TECHNOLOGY Sep. 2007
里氏木霉木聚糖酶基因 XYN 2的克隆与表达
王向明 1 ,欧阳嘉 2 ,严明 1 ,许琳 1
(1. 南京工业大学制药与生命科学学院, 江苏南京 210009;
3 2 2 2. 南京林业大学化学工程学院, 江苏南京 210037)
摘要: 采用 RT PCR ( reverse tra2nscrip tion PCR assay)技术,从里氏木霉 QM9414 mRNA 中扩增出木聚糖酶基因
XYN 2,并与质粒 pYX212相连,构建了组成型表达载体 pYX212 /XYN 2. 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/m in,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.
关键词: RT PCR; Trichoderm a reesei XYN 2基因;酿酒酵母;克隆;表达
中图分类号: Q786 文献标识码: A 文章编号: 1671 - 7627 (2007) 05 - 0082 - 06
C lon ing and expression of the T. reesei xylana se gene XYN 2
WANG Xiang m ing1 , OUYANG J ia2 , YAN M ing1 , XU L in1
(1. College of L ife Science and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China;
2. College of Chem ical Engineering, Nanjing Forestry University, Nanjing 210037, China)
Abstract: X YN 2 gene fragments were essfully amp lified from Trichoderm a reesei strain QM9414 mRNA by RT
PCR ( reverse transcrip tion PCR assay) technology and were ligated to pYX212 vector as the exp ression vector
pYX212 /X YN 2. harom yces cerevisia petent cells were transformed with the binant p lasm id
and began exp ressing. The effects of different carbon sources, temperature, rot