酵母菌3-磷酸甘油脱氢酶基因(gpd1)的克隆及其在高渗胁迫下mRNA转录水平的差异  

Study on Yeast Glycerol-3-Phosphate Dehydrogenase Gene Cloning and Its mRNA Transcriptional Level in the Hyper-osmotic Conditions

作　　者：张凡[1] 马向东[2] 余华顺[2] 姚娟[2] 郭永豪[3] 邢学森[4] 阎淳泰[1] 梁运祥[1] 

机构地区：[1]华中农业大学农业微生物国家重点实验室 [2]湖北安琪酵母股份有限公司博士后工作站,宜昌443003 [3]河南农业大学生物技术与食品科学学院,郑州450002 [4]湖北三峡大学医学院,宜昌443003

出　　处：《食品与发酵工业》2005年第5期15-19,共5页Food and Fermentation Industries

摘　　要：YY、FHS和WFB是生产中常用的酵母菌株。在高渗胁迫下的生长速度依次为FHS、YY、WFB。克隆gpd1基因片段并测序,再与NCBI公布的gpd1序列(X76859)比较,WFB、YY、FHS的同源性分别为99·32%、99·83%、99·83%,YY、FHS菌株的gpd1序列与NCBI上的gpd1序列(X76859)对应的氨基酸序列完全一致,WFB与它们的同源性只有98·5%。用不同浓度的NaCl培养酵母细胞进行渗透胁迫处理,RNA点杂交显示YY、FHS菌株gpd1基因的mRNA表达量在NaCl浓度上升时候有上升的趋势;WFB菌株gpd1基因的mRNA表达量随NaCl浓度的上升逐渐降低。实验结果显示,gpd1基因mRNA转录水平与抗高渗能力相关。YY,FHS and WFB are most commonly used yeast strains. When cultivated in hyper-osmotic conditions, the growth rate of FHS is high, followed by YY and WFB. The results of cloning and sequencing of gpd1 gene from these strains indicate that the nucleotide homologies of the gpd1 gene fragments from strains WFB?YY?FHS 99.32%?99.83%?99.83% compared with the sequences presented on the NCBI website (X76859). As to amino acids, the compared homology are 98.5%?100%, 100% respectively. Disposed with different concentration of NaCl as hyper-osmotic stress, the RNA Dot blotting results show that the mRNA transcriptional levels of gpd1 gene increased for YY, FHS strains whereas decrease for strain WFB as the concentration of NaCl increase. It is found that the yeast ability to endure hyper-osmotic environment could be strengthened by improve the transcriptional mRNA levels of gpd1 gene.

关 键 词：mRNA 转录水平 脱氢酶基因 克隆 NaCl浓度 甘油 磷酸 RNA点杂交 NCBI 氨基酸序列 酵母菌株 生长速度 基因片段 胁迫处理 酵母细胞 同源性 表达量 上升 显示 测序 

分 类 号：Q943[生物学—植物学]