以rDNA为同源重组位点酵母表达鲑鱼降钙素基因多拷贝整合载体的构建  

Construction of a Ribosomal DNA Multi-Copy Integration Vector for Saccharomyces cerevisiae Expressing Salmon Calcitonin

作　　者：姜勇[1] 张学成[1] 孙平楠[2] 陈云松[1] 

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266003 [2]汕头大学医学院,广东汕头515071

出　　处：《中国海洋大学学报（自然科学版）》2009年第3期443-447,共5页Periodical of Ocean University of China

基　　金：国家高技术研究发展计划项目(819204210)资助

摘　　要：根据酵母整合质粒的设计要求,PCR扩增2.2 kb rDNA片段,导入酿酒酵母(Saccharomyces cerevisiae)整合载体pYD1中,设计并合成适合在酵母中表达的鲑鱼降钙素(Salmon Calcitonin),以此为目标基因,构建适合酿酒酵母多拷贝整合的表达载体p-r-sCT。通过载体p-r-sCT引入到酿酒酵母菌株EBY100中,得到酵母重组菌株yAGA2-r-sCT。流式细胞检测结果表明,目标基因鲑鱼降钙素在重组菌株的yAGA2-r-sCT中得到了表达,实现了外源基因在酿酒酵母菌株中的稳定表达。We cloned Salmon Calcitonin （sCT） gene and a 2.2 kb rDNA fragment from Saccharomyces cerevisiae into pYD1 vector, created a new multi-copy intergration plasmid p-r-sCT. This plasmid was gene trans-formed into Saccharomyces cerevisiae EBY100 and obtained a new strain Saccharomyces cerevisiae yAGA2-rsCT. Fluorescent pictures indicated that Salmon Calcitonin was expressed in yAGA2-r-sCT and the quantity of expression was detected by flow cytometry, which means that the sCT gene was integrated into the yast chromosome by a tandem arrangement.

关 键 词：鲑鱼降钙素 酿酒酵母 RDNA 多拷贝整合 流式细胞技术 

分 类 号：Q782[生物学—分子生物学]