簇毛麦中谷胱甘肽硫转移酶基因的克隆与表达分析  

Cloning and Expression Analysis of a Glutathione S-Transferase Gene from Haynaldia villosa

作　　者：何华纲[1] 高安礼[2] 王秀娥[3] 陈佩度[3] 

机构地区：[1]江苏大学食品与生物工程学院,江苏镇江212013 [2]河南大学生命科学学院,河南开封475004 [3]南京农业大学作物遗传与种质创新国家重点实验室,南京210095

出　　处：《河南大学学报（自然科学版）》2008年第4期387-392,共6页Journal of Henan University:Natural Science

基　　金：国家863计划(2001AA222151);国家自然科学基金资助项目(30270828);江苏大学高级专业人才科研启动基金资助项目(05JDG033)

摘　　要：利用抑制性消减杂交(SSH)方法克隆差异表达片段EST639,从簇毛麦cDNA文库中筛选到一个类谷胱甘肽硫转移酶基因(GST)的全长cDNA,命名为HvGSTF.该序列长1 232 bp,具完整的开放阅读框(ORF),编码一条含229个氨基酸残基的多肽,推导其分子量为25.6 kDa,等电点为4.89.该多肽的氨基酸序列与小麦、水稻、大麦、玉米、拟南芥的类GST分别具有83%,63%,61%,52%和47%的相似性.Northern杂交表明,HvGSTF在抗、感病簇毛麦叶片中都为组成型高表达,但在受白粉菌侵染的抗、感病簇毛麦中的表达模式不同,却与活性氧的积累模式存在着一致性,推测HvGSTF在簇毛麦细胞免受活性氧毒害方面起着重要作用.A full-length cDNA of Ф class glutathione S- transferase gene, designated as HvGSTF, was screened from Haynaldia villosa cDNA Library by EST639, a differentially expressed fragment isolated by suppression subtractive hybridization （SSH）. The HvGSTF cDNA was 1 232 bp in length with an open reading frame （ORF） encoding 229 amino acid residues. The deduced mass weight was 25.6 kDa and the pI was 4.89. The deduced protein shares 83%, 63%, 61%, 52% and 47% identity with wheat, rice, barley, maize and Arabidopsis thaliana, respectively. Northern blot showed that HvGSTF gene was constitutively expressed with high level in the leaves of H. villosa and was induced by Erysiphe graminis. There were different expression patterns between resistant Gh21 line and its susceptible mutant M14S line. But the expression pattern coincided well with the reactive oxygen species （ROS） accumulation patterns during invading of pathogens. HvGSTF might play a crucial role in preventing oxidative damage in plant.

关 键 词：簇毛麦 谷胱甘肽硫转移酶基因 抗白粉病相关基因 抑制性消减杂交 

分 类 号：Q78[生物学—分子生物学]