酿酒酵母ARS304及其侧翼序列重组质粒的构建与鉴定  

Construction and Identification of Recombinant Plasmids of Saccharomyces cerevisiae ARS304 and Its Flank Sequence

作　　者：李珺[1] 王馨[1] 张启声[1] 蔡禄[1] 

机构地区：[1]内蒙古科技大学生物工程与技术研究所,内蒙古包头014010

出　　处：《湖北农业科学》2012年第21期4891-4893,4898,共4页Hubei Agricultural Sciences

基　　金：国家自然科学基金(61072129);内蒙古科技大学创新基金(2011NCL005)

摘　　要：采用PCR扩增酿酒酵母(Saccharomyces cerevisiae)Ⅲ号染色体ARS304序列(S1)及以ARS304为核心上下游延伸1 kb的侧翼序列(S2),并将目的片段分别构建到酵母整合载体pRS405中,获得重组质粒pRS1和pRS2。质粒电转化试验发现重组质粒pRS2的转化率大于pRS1,说明ARS304侧翼序列能显著提高ARS304在酿酒酵母中自主复制的能力。ARS304 gene（S1） and its flank sequence（S2） of Saccharomyces cerevisiae were cloned by PCR,and then connected into the shuttle vector pRS405.Recombinant plasmids pRS1 and pRS2 were successfully constructed and used for electroporation of S.cerevisiae.Results showed that the electricity transformation rate of pRS2 was higher than that of pRS1,indicating that flank sequence could improve the autonomously replicating activity of ARS304 in S.cerevisiae.

关 键 词：酿酒酵母(Saccharomyces cerevisiae) 自主复制序列(ARS) 重组质粒 复制起始活性 

分 类 号：Q785[生物学—分子生物学]