转萝卜过氧化物酶基因Rsprx1提高毕赤酵母抗盐性  

Enhancement of Salt Tolerance in Pichia pastoris by Gene Transfer of Radish Peroxidase

作　　者：何静辉[1] 杨静[1] 王丽[1] 李日辉[1] 穆春[1] 贾晓琳[1] 王林嵩[1] 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007

出　　处：《中国生物化学与分子生物学报》2011年第12期1149-1155,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：河南省生物化学与分子生物学重点学科(No.2008169)资助项目~~

摘　　要：为探讨转萝卜过氧化物酶基因(Rsprx1)提高毕赤酵母(Pichia pastoris)抗盐性机理,用不同浓度NaCl处理转基因酵母GSRP25和野生型酵母GS115,检测菌体生长、相对无机盐含量、过氧化物酶活性和同工酶谱及某些抗性基因表达.实验结果表明,在YPD培养条件下,转基因酵母的过氧化物酶活性和菌体生长速率高于野生型酵母,其过氧化氢酶(CTT1)、热休克蛋白(Hsp12)、Rsprx1基因表达和K+/Na+比值均高于野生型.醛脱氢酶(ALD3)的mRNA表达在两者之间没有差异.在BMMY培养条件下,转基因酵母菌体生长速率和过氧化物酶活性显著高于野生型酵母.因此,转基因酵母通过增加过氧化物酶基因表达提高过氧化物酶活性,改变细胞的某些基因表达和无机盐相对含量,从而提高酵母抗盐能力.In order to investigate the effects of peroxidase on the tolerance of yeast against high-salt stress,two strains of Pichia pastoris,including its transgenic and wild type,were treated by different concentrations of NaCl in YPD or BMMY medium.The results showed that the peroxidase activity and growth rate in transgenic strain were higher than those in wild yeast strain with YPD medium.The gene expression of catalase1,heat shock protein12,radish peroxidase1 and the K＋/Na＋ ratio were increased in transgenic yeast strain compared with wild yeast strain.There was no expression difference in alcohol dehydrogenase mRNA between the two strains.The growth rate of transgenic strains and peroxidase activity were much higher than that of wild strain in BMMY medium.Therefore,the peroxidase gene（Rsprx1） of Chinese red radish improves resistance to salinity in yeasts by increasing the expression of peroxidase gene to increase the peroxidase activity,to change the expression of certain genes and the relative content of inorganic salts.

关 键 词：过氧化物酶 基因表达 盐胁迫 毕赤酵母 

分 类 号：Q554.6[生物学—生物化学]