生孢噬纤维菌CMcase基因的分离与表达  

Isolation and Expression of CMcase Gene from Sporocytophaga sp.JL-01

作　　者：刘建[1] 张春玉[2] 李巍[1] 刘东波[2] 

机构地区：[1]长春理工大学生命科学技术学院,长春130022 [2]东北师范大学生命科学学院,长春130024

出　　处：《长春理工大学学报（自然科学版）》2011年第4期122-124,共3页Journal of Changchun University of Science and Technology(Natural Science Edition)

基　　金：国家自然科学基金项目(30670060)

摘　　要：从生孢噬纤维菌中分离完整的基因组DNA,经过PstI限制性内切酶部分酶切后,在T4DNA连接酶作用下,将酶切的片段连接到载体pUC18的PstI位点中,然后转化到E.coli DH5α中,构建了基因组文库,经过筛选获得4.2x103个重组子。经过刚果红平板筛选获得了含有CMCase基因片段的重组子,在E.coli中进行表达,E.coli在37℃培养25~30h,培养液中CMCase活力达到22.8U/mL。In this study,the genomic library of Sporocytophaga sp.JL-01 was constructed.The genom DNA isolated from Sporocytophaga was partially digested with PstI,and then the fragments were ligated to pUC18 vector（PstI） and transformed into E.coli DH5α.4.2x103 recombinants were obtained by means of screening.Recombinant containing a DNA fragment of CMC cellulose（CMCase） achieved by making use of Congo red plate.The CMCase gene was expressed in E.coli.When the E.coli was cultivated at 37℃ for 25~30 hours,the cellulase activity reached 22.8U/mL.

关 键 词：生孢噬纤维菌 CMcase基因 基因组文库 表达 

分 类 号：Q78[生物学—分子生物学]