酿酒酵母细胞cdc50基因敲除及其转录组测序分析  

Knockout and transcriptome sequencing analysis of cdc50 gene in Saccharomyces cerevisiae cells

作　　者：李鑫玉 唐秀琴 李子航 刘佳 王琦 邹伟 LI Xinyu;TANG Xiuqin;LI Zihang;LIU Jia;WANG Qi;ZOU Wei(School of Public Health,Kunming Medical University,Kunming 650500,China;Faculty of Basic Medical Sciences,Kunming Medical University,Kunming 650500,China)

机构地区：[1]昆明医科大学公共卫生学院,云南昆明650500 [2]昆明医科大学基础医学院,云南昆明650500

出　　处：《中国酿造》2024年第4期123-128,共6页China Brewing

基　　金：云南省教育厅科学研究基金项目(2023Y0614)。

摘　　要：为探讨cdc50基因对酿酒酵母(Saccharomyces cerevisiae)细胞功能的影响,该研究以PYM14质粒为模板,采用醋酸锂转化法对野生型酿酒酵母BY4742进行cdc50基因敲除,测定突变菌株Δcdc50生长情况,分别对野生型酵母菌株BY4742和突变菌株Δcdc50进行转录组分析,筛选二者差异表达基因(DEGs),并对其进行基因本体论(GO)、京都基因与基因百科全书(KEGG)富集分析。结果表明,成功构建酿酒酵母cdc50基因缺失菌株Δcdc50,cdc50基因缺失后酵母细胞形态由圆形变为杆状,培养12 h后菌株Δcdc50相对生长率为1.98%,较野生型酵母BY4742菌株显著降低(P<0.05)。与野生型酵母菌株BY4742相比,菌株Δcdc50的DEGs共有581个,其中271个基因上调,310个基因下调。GO富集分析表明,DEGs主要富集在生物学过程中的氧化还原过程等,分子功能中的氧化还原酶活性、辅助因子结合和跨膜转运体活性等。KEGG富集分析表明,DEGs主要富集于糖酵解/糖异生、脂肪酸降解和碳代谢通路等。In order to explore the effect of cdc50 gene on the function of Saccharomyces cerevisiae cells,using PYM14 plasmid as the template,the cdc50 gene in wild type S.cerevicae BY4742 was knocked out by lithium acetate conversion method,and the growth of mutant strainΔcdc50 was measured.Transcriptomic analysis was performed on wild type yeast strain BY4742 and mutant strainΔcdc50 respectively,differential expression genes(DEGs)of them were screened,and the gene ontology(GO)and the Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were carried out.The results showed that the S.cerevisiae cdc50 gene deletion strainΔcdc50 was successfully constructed.After gene cdc50 deletion,the yeast cells morphology changed from round to rod,the relative growth rate of strainΔcdc50 was 1.98%after culture for 12 h,which was significantly lower than that of wild type yeast BY4742(P<0.05).Compared with wild type yeast BY4742,there were 581 DEGs in strainΔcdc50,among them,271 genes were up-regulated and 310 genes were down-regulated.GO enrichment analysis showed that DEGs mainly concentrated in the oxidation-reduction process in biological processes,the oxidoreductase activity,cofactor binding and transmembrane transporter activity in molecular functions.The KEGG enrichment analysis showed that DEGs was mainly concentrated in glycolysis/gluconeogenesis,fatty acid degradation and carbon metabolism pathways.

关 键 词：酿酒酵母 cdc50基因敲除 转录组 差异表达基因 

分 类 号：Q78[生物学—分子生物学] Q93[医药卫生—人体生理学] R33[医药卫生—基础医学]