工业酿酒酵母PGK启动子的克隆与功能分析  

Cloning and Function Analysis of the Promoter of PGK Gene in Saccharomyces cerevisiae

作　　者：阳辛凤[1,2,3] 郭安平[2] 孔华[2] 郭运玲[2] 贺立卡[2] 

机构地区：[1]海南大学农学院,海口570228 [2]中国热带农业科学院热带生物技术研究所,海口571101 [3]中国热带农业科学院分析测试中心,海口571101

出　　处：《中国农学通报》2012年第6期178-182,共5页Chinese Agricultural Science Bulletin

基　　金：国家"863"重点课题"生物质转化微生物资源的开发利用"(No.2007AA021307);转基因生物新品种培育重大专项课题"转基因生物安全监测技术--转基因作物南繁环境影响监测"(No.2008ZX08012-04);国家"973"计划项目"木薯重要基因的功能验证与分析"(No.2010CB126605)

摘　　要：为了获得适用于构建工业酿酒酵母整合型表达载体的组成型启动子,以工业酿酒酵母南阳K基因组DNA为模板,采用PCR方法扩增得到了磷酸甘油酸激酶基因起始密码前的启动子片段2个,其中长片段781bp,命名为PGK1(GenBank Acession No.FJ415226)。NCBIBlast软件分析结果表明,PGK1核苷酸序列与酿酒酵母染色体Ⅲ上PGK启动子(GenBank Acession No.X59720)相似性为99%,序列中含有基因表达所需的基本调控元件TATA-box和CAAT-box等。功能分析表明PGK1能驱动整合在工业酿酒酵母基因组上的外源基因葡萄糖淀粉酶基因的表达。综上表明成功克隆得到PGK1启动子,为工业酿酒酵母表达载体的构建奠定了基础。The aim was to obtain a constitutive promoter for construction of expression vector of industrial Saccharomyces cerevisiae.Applying S.cerevisiae NYK genomic DNA as template,two fragments upstream the initiate codes gene were obtained by PCR.The long fragment was 781 bp and designated PGK1（GenBank Accession No.FJ415226）.Sequence analysis by NCBI Blast showed that,the nucleotide sequence of PGK1 was 99% homologous to PGK promoter located in chromosome Ⅲ from S.cerevisiae（GenBank Accession No X59720）.TATA box,CAAT-box and other regulatory elements for gene expression were detected in the nucleotide sequence of PGK1.Functional analysis indicated that,PGK1 could initiate the expression of the foreign glucoamylase gene（ga1） integrated in industrial S.cerevisiae.The results showed that,promoter PGK1 was obtained which provided experimental base for construction of expression vector of industrial S.cerevisiae.

关 键 词：酿酒酵母 PGK启动子 克隆 序列分析 

分 类 号：Q812[生物学—生物工程]