朝鲜碱茅过氧化氢酶基因(PuCAT)的克隆及表达分析  被引量:2

Molecular Cloning and Expression Analysis of Catalase(PuCAT)Gene in Puccinellia chinampoensis

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作  者:任伟[1,2] 徐博[3] 耿慧[1] 王志锋[1] 徐安凯[1] 

机构地区:[1]吉林省农业科学院畜牧科学分院,吉林公主岭136100 [2]中国科学院水土保持与生态环境研究中心,陕西杨凌712100 [3]吉林农业大学,吉林长春130000

出  处:《北方园艺》2014年第20期90-95,共6页Northern Horticulture

基  金:现代农业产业技术体系建设专项资金资助项目(cars-35-02)

摘  要:以朝鲜碱茅(Puccinellia chinampoensis)茎叶组织提取的RNA为模板,根据已报道的CAT基因同源序列设计引物,通过RT-PCR法扩增出1个CAT基因的cDNA序列并与其它植物CAT基因进行同源性比对。结果表明:该基因全长1 487bp,是一个完整开放阅读框,编码含492个氨基酸的蛋白,Genebank登陆号为:HM230827,命名PuCAT;朝鲜碱茅CAT基因的核苷酸和氨基酸序列与大麦、小麦的同源性最高;并对其信号肽、疏水性、跨膜结构、二级结构和主要功能域做了预测。半定量RT-PCR结果表明,PuCAT基因在4种胁迫处理条件下,有着不同的表达规律,总体而言,在朝鲜碱茅的根部和叶片均可以表达,但是在叶片中的表达量要高于在根中的表达。The full length cDNA sequence of CAT gene was cloned from Puccinellia chinampoensis leaves using RT-PCR method,the primers were designed according to the homologous CAT gene sequences of other plant species. The results showed that the nucleotide sequence of the gene was 1 487 bp,containing a complete open reading frame and encoding 492 amino acids, Genebank: HM230827. Nucleotide and amino acid sequence analysis revealed that PuCAT shared high identity with the orthologs from Triticum aestivum and Hordeum vulgare. And its signal peptide, hydrophobicity/ hydrophilic,trans-membrane domain, secondary structure and main functional domains were predicted. Semi-quantitative RT-PCR analysis showed that PuCAT expressed in different tissues,but the expression in root was lower,and in leaf much higher. Various elevated levels of PuCAT expression had been detected when exposed to 4 different stress experimental treatments ,and the results were not the same.

关 键 词:朝鲜碱茅 过氧化氢酶 基因克隆 

分 类 号:Q785[生物学—分子生物学]

 

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