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酿酒酵母GPD2基因沉默表达载体的构建

发表时间:2016-02-15  浏览量:3097  下载量:1139
全部作者: 王斌,苏秋琼,冯佳乐,叶晓丹,薛婷,陈由强
作者单位: 福建师范大学生命科学学院;福建师范大学福建省发育与神经生物学重点实验室
摘 要: 根据5′-UTR的二级结构能够阻碍基因转录的原理,以PCR方法为基础获得酿酒酵母(Saccharomyces cerevisiae)GPD2基因5′-UTR的反义RNA片段SGPD2基因沉默片段。通过限制性内切酶的酶切和T4连接酶的连接,将SGPD2基因片段与本实验室保存酿酒酵母表达载体pUPST构建GPD2基因的沉默表达载体pUPT-SGPD2,为下一步构建能够提高乙醇产量的酿酒酵母GPD2沉默菌株的研究奠定基础。
关 键 词: 分子生物学;反义RNA;3-磷酸甘油脱氢酶;GPD2基因;酿酒酵母
Title: Constructing GPD2 gene silencing expression vector for Saccharomyces cerevisiae
Author: WANG Bin, SU Qiuqiong, FENG Jiale, YE Xiaodan, XUE Ting, CHEN Youqiang
Organization: College of Life and Science, Fujian Normal University; Key Laboratory of Developmental and Neural Biology, Fujian Normal University
Abstract: According to the principle of the 5′-UTR of secondary structure hindering gene transcription, we constructed antisense RNA gene silencing fragment SGPD2 for Saccharomyces cerevisiae GPD2 gene 5'-UTR based on the method of PCR.Expression vector pUPST of S. cerevisiae was preserved in our laboratory. We built GPD2 gene silence express carrier pUPT-SGPD2, with SGPD2 gene fragments and S. cerevisiae expression vector pUPST by restriction enzymes enzyme digestion and T4 ligase connection. The construction of pUPT-SGPD2 gene silence express vector provided the foundation for the ethanol production of S. cerevisiae GPD2 silence strains research.
Key words: molecular biology; antisense RNA; 3-phosphate glycerol dehydrogenase; GPD2 gene; Saccharomyces cerevisiae
发表期数: 2016年2月第3期
引用格式: 王斌,苏秋琼,冯佳乐,等. 酿酒酵母GPD2基因沉默表达载体的构建[J]. 中国科技论文在线精品论文,2016,9(3):213-218.
 
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