酵母α-半乳糖苷酶基因在大肠杆菌中的表达  

Expression of Alpha-galactosidase Gene MEL1 in Escherichia Coli

作　　者：周双德 张学文[2] 唐香山[2] 

机构地区：[1]湖南省农业生物工程研究所,湖南长沙410128 [2]湖南农业大学生命科学与技术学院,湖南长沙410128

出　　处：《长沙理工大学学报（自然科学版）》2005年第1期92-95,共4页Journal of Changsha University of Science and Technology:Natural Science

基　　金：湖南省科技厅中青年基金项目(01JZY2099)

摘　　要：酵母编码α-半乳糖苷酶的基因MEL1被扩增并克隆到表达载体pRSET中.重组质粒pRSET Gal转化至相应的受体菌BL21(DE3)PlysS,阳性克隆经液体培养和IPTG诱导表达.通过SDS PAGE分析,在50kD处有一目的分子大小的亮带.裂解细菌后经X α Gal的显色底物反应,液体变蓝.试验表明:α-半乳糖苷酶基因MEL1在大肠杆菌中得到了表达,糖基化对于维持此酶的生物活性不是必须的.MEL1 gene was amplified in saccharomyces cerevisiae AH109 with PCR and recombined into E.coli expression vector pRSET. Recombinant plasmid pRSET-Gal was obtained and transformed into host E.coli BL21(DE3)PlysS. Transformed strain was cultured in liquid LB and induced with IPTG.There is a bright band about 50kD detected by SDS-PAGE analysis. X-α-Gal was added into lysed cultures;blue reaction indicated alpha-galactosidase was expressed in E.coli; glycosylation was not needed to keep activity of alpha-galactosidase.

关 键 词：半乳糖苷酶基因 大肠杆菌 酵母 SDS-PAGE分析 α-半乳糖苷酶 表达载体 重组质粒 诱导表达 IPTG 液体培养 阳性克隆 分子大小 生物活性 受体菌 糖基化 底物 细菌 

分 类 号：Q78[生物学—分子生物学] TS261.11[轻工技术与工程—发酵工程]