酿酒酵母PMT6基因缺失对细胞寿命的影响  

Effect of PMT6 gene deficiency on the cell lifespan of Saccharomyces cerevisiae

作　　者：崔红晶 周涛 赵炜 宋浩昌 袁源 郑慧玲 刘新光 

机构地区：[1]广东医科大学衰老研究所广东省医学分子诊断重点实验室东莞市衰老与抗衰老重点实验室,广东东莞523808

出　　处：《生物技术》2017年第6期576-579,544,共5页Biotechnology

基　　金：国家自然科学基金-青年基金项目("细胞自噬介导PMT1基因调节酵母细胞寿命的作用机制研究";No.31701050);广东省医学科学技术研究基金项目("内质网应激介导PMT1基因缺失延长酵母复制性寿命的机制研究";No.A2016257);东莞市医疗卫生科技计划一般项目("细胞自噬在PMT1基因缺失延长酵母复制性寿命中的作用和机制";No.2016105101289)

摘　　要：[目的]构建蛋白质O-甘露糖转移酶(Protein O-mannosyltransferase)-6基因缺失酵母菌株,研究其对细胞寿命的影响。[方法]以pRS306质粒为模板,采用一步基因置换方法构建PMT6基因缺失菌株(pmt6Δ);光学显微镜下分离和计数酵母母细胞产生子细胞的数目,检测酵母细胞的复制性寿命;在热量限制条件下(含0.5%葡萄糖培养基),检测酵母细胞分裂增殖速度。[结果]成功构建pmt6Δ菌株;与对照组酵母细胞的复制性寿命(25.2代)比较,pmt6Δ菌株的寿命(25.4代)无明显变化(P>0.05);在热量限制条件下,pmt6Δ菌株的生长曲线低平,细胞分裂增殖减慢。[结论]PMT6基因不参与酵母细胞复制性寿命的调控,而热量限制影响PMT6基因缺失细胞的分裂增殖能力。[Objective] To study the effect of the protein O-mannosyltransferases 6（PMT6） gene deficiency on the cell proliferation of yeast.[Methods] PMT6 deficiency strain（pmt6Δ） was constructed through polymerase chain reaction（PCR）-mediated one-step gene disruption.The replicative lifespan was counted by separating daughter cells from mother cells using a manual micromanipulator equipped with a fiber-optic needle under the microscope.Cell proliferation of pmt6Δ strain was assessed using the full automatic growth curve analyzer under calorie restriction condition（containing 0.5% glucose in medium）.[Results] PMT6 deleted strain（pmt6Δ） was successfully constructed.Mean replicative lifespan of the pmt6Δ yeast strain（25.4 generations） exhibited no significant difference,compared with that of the control strain（25.2 generations）（p 〉0.05）,and the growth curves of pmt6Δ strain showed the slow growth,compared with of that of the control strain under calorie restriction condition.[Conclusion] PMT6 deficiency exerted no evident effect on the replicative lifespan of yeast cells,and while calorie restriction affected the cell proliferation of pmt6Δ yeast strain.

关 键 词：酿酒酵母 热量限制 复制性寿命 细胞增殖 

分 类 号：Q291[生物学—细胞生物学]