芽殖酵母Sik1蛋白在纺锤体定位中的功能  

Functional study of the Saccharomyces cerevisiae Sik1 protein in spindle orientation

作　　者：唐仙英 庞文颖 海力 滕雪 张楠 朱顺 TANG Xianying;PANG Wenying;HAI Li;TENG Xue;ZHANG Nan;ZHU Shun(Key Laboratory of Hubei Province for the Protection and Utilization of Special Plant Germplasm in Wuling Mountain Area & Key Lab for Biotechnology of State Ethnic Affairs Commission &College of Life Sciences, South-Central University for Nationalities, Wuhan 430074, China)

机构地区：[1]中南民族大学生命科学学院&武陵山区特色资源植物种质保护与利用湖北省重点实验室&生物技术国家民委重点实验室,武汉430074

出　　处：《中南民族大学学报（自然科学版）》2021年第3期252-257,共6页Journal of South-Central University for Nationalities：Natural Science Edition

基　　金：国家自然科学基金资助项目(31401155);中央高校基本科研业务费专项资金资助项目(CZY18021)。

摘　　要：为了探究芽殖酵母蛋白Sik1在有丝分裂纺锤体定位中的功能,利用同源重组技术将SIK1的内源启动子替换成GAL1启动子,得到基因组位点过表达SIK1的菌株,同时使菌株表达GFP-Tub1,以便观察纺锤体;接着在12℃低温条件下培养细胞以促进双核细胞的产生,细胞经乙醇固定、DAPI染色后在荧光显微镜下观察细胞核分裂及纺锤体定位;最后检测SIK1过表达对细胞生存能力的影响.结果表明:过表达Sik1会干扰纺锤体定位,但对细胞核迁移没有明显影响.在16℃低温胁迫条件下,过表达Sik1降低了纺锤体定位突变体kar9Δ的生长活力却部分恢复了num1Δ细胞的生长,说明Sik1可能在Dynein通路中起作用.To explore the function of Saccharomyces cerevisiae Sik1 protein in spindle orientation,the endogenous promoter of SIK1 was replaced with the GAL1 promoter through homologous recombination approach to obtain the strain that overexpresses SIK1 in the chromosomal locus.The strain was also constructed to express GFP-Tub1 to observe spindles.Next,cells were incubated at 12℃to facilitate the production of bi-nucleated cells.Cells were fixed with ethanol,stained with DAPI,and observed with fluorescence microscope to examine nuclear division and spindle orientation.Finally,the effect of overexpressing Sik1 on cell viability was examined.The results showed that overexpressing Sik1 disturbed spindle orientation,but had no evident negative effect on nuclear migration.Under the stress of low temperature 16℃,overexpression of Sik1 reduced the viability of kar9Δ,a spindle orientation mutant,but recovered the viability of num1Δcells partially,suggesting that Sik1 might play a role in the Dynein pathway.

关 键 词：芽殖酵母 纺锤体定位 Sik1 Num1 核迁移 

分 类 号：Q253[生物学—细胞生物学]