拟南芥WRI1转录因子对酿酒酵母脂肪酸合成的影响  

Effects of the transcription factor WRI1 from Arabidopsis thaliana on the fatty acid synthesis in Saccharomyces cerevisiae

作　　者：闵文莉 曹喜涛[1,2] 季更生[1,2] 屠洁[1,2] 李强 陈欣[1,2] 张国政[1,2] 武国华 MIN Wenli;CAO Xitao;JI Gengsheng;TU Jie;LI Qiang;CHEN Xin;ZHANG Guozheng;WU Guohua(School of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang 212018,China;The Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang 212018,China)

机构地区：[1]江苏科技大学生物技术学院,镇江212018 [2]中国农业科学院蚕业研究所,镇江212018

出　　处：《江苏科技大学学报（自然科学版）》2018年第6期873-877,共5页Journal of Jiangsu University of Science and Technology:Natural Science Edition

基　　金：江苏科技大学博士启动基金项目(1732921203);江苏科技大学生物技术学院蚕业研究所科研基金项目(2014SY10;2015SY02)

摘　　要：转录因子是植物体内一类应答环境胁迫所产生的蛋白,通过激活或抑制转录的方式发挥调控作用. WRI1转录因子是油脂合成的关键因子,主要在脂肪酸合成与糖酵解后期发挥调控作用.为了获得高产脂肪酸的酵母菌株,以拟南芥c DNA为模板,PCR扩增得到编码拟南芥WRI1转录因子的基因,经过Spe I/Bam H I双酶切后连接到表达载体上,得到重组质粒p416-Atwri1.将重组质粒用PEG/Li Ac化学转化法转入酿酒酵母BY4742中,通过营养缺陷型培养基进行筛选,挑取阳性菌落并进行摇瓶发酵,采用GC-MS对脂肪酸进行定性和定量分析.结果表明,Atwri1基因在酵母中的表达改变了脂肪酸的组成及含量,与空质粒酵母相比,重组酵母中长链脂肪酸的总含量增加了44. 8%,为以后酿酒酵母生产脂肪酸实现工业化打下基础.Plant transcription factors are a kind of proteins produced in response to the environmental stress,which exhibit the regulatory function by activating or inhibiting the gene transcription.The WRI1 transcription factor is a key factor to regulate the fatty acid biosynthesis and the late glycolysis.To obtain the yeast strain with high productivity of fatty acids,the WRI1 gene from Arabidopsis thaliana was amplified by PCR,and digested by two restriction endonucleases Spe I and BamH I and ligated to the p416 plasmid for construction of recombinant expression plasmid p416-Atwri1.Then p416-Atwri1 was transformed into Saccharomyces cerevisiae BY4742 by PEG/LiAc method.The positive colonies were screened through auxotrophic medium and then fermented under shake-flask conditions.Gas Chromatograph-Mass Spectrometer-computer was utilized to analyze the compositions and concentrations of fatty acids.The results revealed that the overexpression of Atwri1 gene changed both compositions and concentrations of fatty acids.Compared with the strain harboring the empty p416 plasmid,the long-chain fatty acids in the strain harboring p416-Atwri1 were increased by 44.8%.The engineered yeast provides a foundation for industrial production of fatty acids.

关 键 词：拟南芥 WRI1转录因子 酿酒酵母 脂肪酸 

分 类 号：U661.4[交通运输工程—船舶及航道工程]