ScINO1基因克隆及酵母多基因多拷贝整合表达载体的构建  

Cloning of ScINO1 Gene and Construction of Yeast Multi-copy Integration Expression Vector

作　　者：黄贞杰[1,2,3] 陈玲[1,2,3] 张积森[1,2,3] 叶冰莹[1,2,3] 陈由强[1,2,3] 陈如凯 

机构地区：[1]福建师范大学生命科学学院,福建福州350108 [2]农业部福建甘蔗生物学与遗传育种重点实验室,福建福州350108 [3]福建省发育与神经生物学重点实验室,福建福州350108

出　　处：《福建师范大学学报（自然科学版）》2012年第6期100-105,共6页Journal of Fujian Normal University：Natural Science Edition

基　　金：现代农业产业技术体系建设专项资金资助项目(CARS-20-4-4)

摘　　要：利用PCR技术,以酿酒酵母S288c的基因组DNA为模板,扩增酿酒酵母肌醇-1-磷酸合成酶基因(ScINO1)包含1 602bp的开放读码框,克隆后经PCR和酶切反应进行了鉴定,测序结果与GenBank上已登录的ScINO1序列完全一致.对PUG6载体进行改造,构建了rDNA介导的多拷贝整合表达载体pURH,并将ScINO1基因连入载体pURH,获得ScINO1基因超表达载体pURIH.为下一步的微生物发酵法产肌醇和获得能够耐高乙醇的酵母工程菌株的研究奠定基础.The inositol-l-phosphate synthase encoding gene （SclNO1） was amplified from the genomic DNA of Saccharornyces cerevisiae S288c by PCR. The length of SclN01 gene was 1 602 bp. The Blast results showed that SclN01 gene sequence was consistent with sequence of S. cerevisiae S288c in GenBank. The digested products of PGK, CYC1 and a rDNA fragment were inserted into S. cerevisiae shuttle plasmid PUG6 to generate rDNA mediated multi-copy integrated expression vectors pURH. Then, to obtain multi-copy inte- grated expression vectors pURIH, the SclN01 gene was inserted into pURH. The yeast mul- ti-copy integration expression vector was further verified through double restriction enzyme digestion. The construction of yeast multi-copy integration expression vector was provided with basic material for further study in genetic engineering of synthesis inositol and high ethanol tolerance in yeast.

关 键 词：ScINO1基因 多拷贝整合 载体构建 酿酒酵母 

分 类 号：Q946[生物学—植物学]