紫苏PfDGAT1酵母功能互补分析  

Functional complementation analysis of H1246 yeast mutation by PfDGAT1 gene from Perilla frutescens

作　　者：王计平[1] 周雅莉[1] 任文燕 安茜[1] 段露露 郝月茹 李润植[1] Wang Jiping;Zhou Yali;Ren Wenyan;An Xi;Duan Lulu;Hao Yueru;Li Runzhi(College of Agriculture,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学农学院

出　　处：《山西农业大学学报（自然科学版）》2019年第5期10-15,共6页Journal of Shanxi Agricultural University(Natural Science Edition)

基　　金：国家青年科学基金(31201266);山西省重点研发计划项目(201803D221005-9);山西省晋中市科技重点研发计划项目(Y172007-5)

摘　　要：[目的]本试验将已克隆的紫苏种子中高表达二酰甘油酰基转移酶(DGAT)基因 PfDGAT 1 转化至酿酒酵母突变型菌株H1246进行酵母互补功能验证,研究二酰甘油酰基转移酶是否具有催化油脂合成的作用。[方法]通过构建紫苏 DGAT 基因的酵母表达载体pYES2.0- PfDGAT 1 并转化至酿酒酵母突变型菌株H1246,用尼罗红对转基因酵母细胞染色,检测转基因酵母中是否能合成油体;利用薄层层析(TLC)试验检测转基因酵母中是否存在三酰甘油(TAG)。[结果]转pYES2.0- PfDGAT 1 载体的突变型酵母菌株H1246能够检测到有油体存在,而转pYES2.0空载体的突变型酵母菌株H1246中没有检测到油体,说明紫苏 PfDGAT 1 基因的表达恢复了酿酒酵母突变型菌株H1246油体缺陷的表型,可以催化TAG合成。[结论] PfDGAT 1 基因在紫苏种子油脂合成积累过程中发挥重要作用,该研究结果为深入解析 PfDGAT 1 基因在紫苏油脂合成途径中的功能奠定了基础。[Objective]In the previous study,the diacylglycerolacyltransferase (DGAT) gene PfDGAT 1 has been cloned from perilla seeds and highly expressed.In this study,yeast Saccharomyces cerevisiae mutant strain H1246 was used to test the function of DGAT for catalyzing oil synthesis by yeast functional complementation experiments.[Methods]The yeast expression vector pYES2.0- PfDGAT 1 of PfDGAT 1 gene was successfully constructed by double digestion,and the recombinant plasmid was transformed into the yeast mutant strain H1246.The transgenic yeast cells were stained with Nile Red to detect whether oil body could be synthesized in the transgenic yeast.TLC test was used to detect TAG presented in thransgenic yeast.[Results] Oil bodies were able to be detected in the mutant strain H1246 transfected with pYES2.0- PfDGAT 1 ,but not in the mutant strain H1246 transfected with pYES2.0 empty vector,whichindicated that the oil body defectingin Saccharomyces cerevisiae mutant strain H1246was restored by the expression of PfDGAT 1 gene.The diacylglycerolacyltransferase could catalyze TAG synthesis.[Conclusion] The PfPDAT1 gene plays an important role in the synthesis and accumulation of perilla seed oil.These results lay the foundation for the further study of the function of PfDGAT 1 gene in the fatty acid metabolism pathway of perilla.

关 键 词：紫苏 二酰甘油酰基转移酶 酵母功能互补 油脂合成 

分 类 号：S565.8[农业科学—作物学]