盐穗木谷胱甘肽转硫酶基因的克隆和表达分析  

Molecular Cloning and Expression Analysis of Glutathione S-Transsulfurase Gene in Halophyte Halostachys caspica

作　　者：地丽热巴.地里夏提 张霞[1] 庄淑珍[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院/新疆生物资源与基因工程重点实验室,乌鲁木齐830046

出　　处：《新疆农业科学》2014年第2期318-324,共7页Xinjiang Agricultural Sciences

基　　金："973"计划前期研究专项(2012CB722204);新疆生物资源基因工程重点实验室开放课题(XJDX0201-201205)

摘　　要：[目的]植物抗氧化酶在逆境条件下会产生胁迫响应,克隆盐穗木（Halostachys caspica）谷胱甘肽转硫酶（glutathione s-transferase,GST）基因并进行表达分析.[方法]根据盐穗木盐胁迫抑制差减文库获得的部分EST序列,通过RT-PCR和RACE技术,从盐穗木中获得盐穗木谷胱甘肽转硫酶基因（HcGST）的全长cD-NA序列.[结果]HcGST基因全长1 009bp,其中3＇-非翻译区为250 bp,5＇-非翻译区为85 bp,开放阅读框为675 bp,编码224个氨基酸.序列比对发现HcGST与其他植物GST蛋白序列同源性高达83％～98％.实时定量PCR检测结果显示,盐胁迫和过氧化氢处理均使HcGST的基因表达上调.[结论]HcGST与其他植物谷胱甘肽转硫酶相比具有高度同源性,对盐胁迫和氧化胁迫均有上调响应.[Objective] Plant antioxidant enzymes under stress conditions produce stress responses,In this study,glutathione S-transferase gene（HcGST） of Halostachys caspica was cloned and the expression analysis of HcGST was analyzed.[Method] Based on the partial EST sequence from SSH libraries in Halostachys caspica under salt stress,the full sequence of glutathione s-transferase gene cDNA was obtained with RT-PCR and RACE technology.[Result] the full length of HcGST was 1,009 bp with 250 bp of 3＇untranslated region （UTR）,85 bp of 5＇UTR,as well as 675 bp open reading frame（ORF） encoding 224 aa.Sequence blast results showed that HcGST displayed the high homologous from other species,about 83％ ～ 98％.The real-time PCR indicated that the transcriptional level of HcGST salt stress and H2O2 treatment were up-regulation.It also demonstrated HcGST belonged to a stress responsive gene.[Conclusion] HcGST was highly homologous compared with other plant glutathione S-transferase,and HcGST expressions were up-regulation responses to salt stress and oxidative stress.

关 键 词：盐穗木 谷胱甘肽转硫酶 盐胁迫 过氧化氢胁迫 表达分析 

分 类 号：S188[农业科学—农业基础科学]