α-半乳糖苷酶pYD1-84411质粒构建  

Construction of α-galactosidase pYD1-84411 Recombinant Plasmid

作　　者：李楠[1] 董墨思 李润国[2] 林利萍[2] 郑立博[2] 李苏红[2] 

机构地区：[1]沈阳师范大学化学与生命科学学院,辽宁沈阳110034 [2]沈阳师范大学粮食学院,辽宁沈阳110034

出　　处：《食品研究与开发》2013年第24期232-235,共4页Food Research and Development

基　　金：国家自然科学基金面上项目(31271819)

摘　　要：以水稻α-半乳糖苷酶重组质粒pET32a+-84411为模板,通过PCR反应条件的筛选,琼脂糖凝胶电泳检测,扩增出条带清晰、单一的水稻α-半乳糖苷酶(BAC84411.1)目的基因,通过双限制性内切酶的酶切插入到酵母表面展示载体pYD1中,此目的基因与载体以摩尔数比为4∶1的比例进行连接。并以将重组质粒转化到大肠杆菌JM109中,并提取到了阳性重组子质粒,以备转化到酵母表达菌株EBY100中,诱导其表达α-半乳糖苷酶。Riceα-galactosidase (BAC84411) recombinant plasmid pET32a+-84411 as template, PCR amplifica-tion conditions and the reaction system were screened. The amplified fragment were detected by agarose gel electrophoresis, a clear and single band of the target gene was obtained. And the gene was inserted into the yeast surface display vector pYD1 by double restriction endonuclease. the insert and the vector ligased by moles ratio of 4∶1. Then the ligation mixture was transformed into E.coli JM109, and the positive recombinant plasmid was identified. The obtained pYD1-84411plasmid would be used for inducing α-Galactosidase expression in yeast strains EBY100.

关 键 词：Α-半乳糖苷酶 酵母表面展示 重组质粒 

分 类 号：Q93[生物学—微生物学]