摘要
为研究蛋白质O-甘露糖转移酶-1(Protein O-mannosyltransferase-1,Pmt1p)与Pmt5p基因对酵母细胞寿命的影响,采用一步基因置换法构建PMT5基因缺失菌株(pmt5Δ),在此基础上,缺失PMT1基因,构建PMT1和PMT5双基因缺失菌株(pmt1Δpmt5Δ)。显微镜下分离和计数酵母子细胞的数目,统计菌株的复制性寿命;检测细胞吸光度值来评价细胞分裂增殖速度。结果发现,与对照组酵母细胞的平均复制性寿命(25代)比较,pmt5Δ菌株(26代)的寿命无明显变化(P> 0. 05),而pmt1Δpmt5Δ菌株(21代)的寿命明显缩短(P <0. 01);热量限制条件下,与对照组酵母细胞比较,pmt5Δ菌株的生长曲线无明显变化,而pmt1Δpmt5Δ菌株的生长曲线低平,细胞分裂增殖减慢。结果表明,PMT1与PMT5双基因缺失明显缩短酵母细胞的寿命,机制可能与细胞的增殖活力下降有关。
In order to investigate the roles of protein O -mannosyltransferase-1 （Pmt1p） and protein O -mannosyltransferase-1 （Pmt5p） on yeast cells lifespan, PMT5 deficiency strain （ pmt5Δ ）, and PMT1 and PMT5 double-gene deficiency strain （ pmt1Δpmt5Δ ） were constructed adopting one-step replacement method in this study. The replication lifespan were then evaluated by separating daughter cells from mother cells using a manual micromanipulator equipped with a fiber-optic needle under the microscope. Cell proliferation of these yeast strains were further determined utilizing the full automatic growth curve analyzer under calorie restriction condition （Yeast medium containing 0.5% glucose）. The results showed that, the mean lifespan of pmt5Δ strain exhibited no significant difference （ P 〉0.05）, while the mean lifespan of pmt1Δ pmt5Δ strain was significantly shortened （ P 〈0.01） as compared with that of the wild-type strain. In addition, PMT5 deficiency conferred no notably difference to the growth of the yeast cells, while both genes deficiency endowed the yeast cells with slower growth rate under calorie restriction condition. The data suggested that PMT1 and PMT5 double-gene deficiency shortened the yeast cells lifespan, the mechanism was probably due to the declined capability of cell proliferation.
作者
崔红晶
周涛
赵炜
宋浩昌
袁源
郑慧玲
刘新光
CUI Hong-jing;ZHOU Tao;ZHAO Wei;SONG Hao-chang;YUAN Yuan;ZHENG Hui-ling;LIU Xin-guang(Inst.of Aging Res;Guangdong Prov.Key Lab.of Med.Mol.Diagnostics;Aging and Anti-Aging Key Lab.of Dongguan,Guangdong Med.Uni.,Dongguan 523808)
出处
《微生物学杂志》
CAS
CSCD
2018年第5期34-38,共5页
Journal of Microbiology
基金
国家自然科学基金项目(31701050
81671399)
广东省医学科学技术研究基金项目(A2016257
B2014299)
东莞市医疗卫生科技计划一般项目(2016105101289)
东莞市国际科技合作(含港澳台)项目(201650812001)
关键词
酿酒酵母
复制性寿命
热量限制
细胞增殖
Saccharomyces cerevisiae
replication lifespan
calorie restriction
cell proliferation
