匍枝根霉内切葡聚糖酶基因在毕赤酵母中的克隆及表达  

Cloning and expression of endoglucanase gene from a cDNA library of rhizopus stolonifer in P. pastoris

作　　者：张莹莹[1] 汤斌[1] 

机构地区：[1]安徽工程大学微生物发酵安徽省工程技术研究中心,安徽芜湖241000

出　　处：《安徽工程大学学报》2013年第2期31-34,共4页Journal of Anhui Polytechnic University

基　　金：国家自然科学基金资助项目(31270315)

摘　　要：从匍枝根霉TP-02的cDNA文库中克隆得到其内切葡聚糖酶基因,并在毕赤酵母中进行表达.该内切葡聚糖酶基因大小为954bp,将其与pPIC9K质粒经EcoRⅠ和NotⅠ双酶切后,利用T4连接酶进行连接.将线性化后的重组质粒转化毕赤酵母GS115,并利用MD平板和刚果红平板进行筛选.阳性克隆经甲醇诱导培养84h后,产生的内切葡聚糖酶酶活力达到峰值为1.754IU/mL.A novel endoglucanase gene was cloned from a cDNA library of the filamentous fungus Rhizopus stolonifer vat. reflexus TP-02 and expressed in P. pastoris. Sequence analysis of the gene revealed an open reading frame of 954 bp. The endoglucanase gene eg2 and the pPIC9K plasmid were digested by EcoRI and Not I, and linked with T4 DNA ligase. The recombinant plasmid was linearized and transformed into P. pastoris, and the positive clones were screened by the MD plate and the Congo Plate. The maximum endoglucanase activity of the recombinant induced by methanol is 1. 754 IU/mL observed at 84 h during the ferment process.

关 键 词：匍枝根霉 内切葡聚糖酶 毕赤酵母 表达 

分 类 号：Q93[生物学—微生物学]