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

[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.