酿酒酵母PMT1与PMT3双基因缺失对细胞寿命的影响

Effects of PMT1 and PMT3 Double-gene Deficiency on the Lifespan of Saccharomyces cerevisiae

研究蛋白质O-甘露糖转移酶(Protein O-mannosyltransferase,PMT)家族中PMT3基因缺失,以及PMT1与PMT3双基因缺失对酵母细胞复制性寿命的影响。采用一步基因置换法构建PMT3基因缺失酵母菌株(pmt3Δ),基于基因同源重组的原理,构建PMT1和PMT3双基因缺失酵母菌株(pmt1Δpmt3Δ);显微镜下分离和计数酵母子细胞的数目,统计酵母细胞复制性寿命。与野生型酵母细胞的平均复制性寿命(26代)比较,pmt3Δ菌株(24代,P>0.05)和pmt1Δpmt3Δ菌株(25代,P>0.05)的平均寿命均无明显变化,差异无统计学意义。酵母PMT3单基因缺失,以及PMT1和PMT3双基因缺失均不影响细胞的复制性寿命。

We herein aimed to explore the effects of the PMT3 of protein O-mannosyltransferase（PMT）single-gene deficiency,and PMT1 and PMT3 double-gene deficiency on the replicative lifespan of Saccharomyces cerevisiae. PMT3 deficiency strain（pmt3Δ）wasconstructed through polymerase chain reaction（PCR）-mediated one-step gene disruption,and PMT1 and PMT3 double-gene deficiencystrain（pmt1Δpmt3Δ）was prepared based on homologous recombination. The replicative lifespan of yeast cells were counted by separatingdaughter cells from mother cells using a manual micromanipulator equipped with a fiber-optic needle under the microscope. The replicativelifespan of the pmt3Δ yeast strain（24 generations）and pmt1Δpmt3Δ strain（25 generations,P 0.05）exhibited no significant difference,compared with that of wild-type yeast strain（26 generations）（P 0.05）,i.e.,there was no statistical significance among the differences.Conclusively,PMT3 single-gene deficiency and PMT1 and PMT3 double-gene deficiency exert no evident effects on the lifespan of yeast cells.