湖南省科技厅重点项目(2010TP4004-1,2010FJ2013,2012FJ4063);国家科技支撑计划(2012BAD20B05-04)
本研究根据苎麻转录组测序结果中的抗坏血酸过氧化物酶(APX)的基因片段,利用RT-PCR结合RACE方法从苎麻(Boehmeria nivea L.)中克隆到一个APX基因的全长cDNA,命名为BnAPX1。BnAPX1 cDNA全长1 201 bp,开放阅读框(ORF)为870 bp,推测其编码一个含289个氨基酸序列的多肽。生物信息学分析表明,BnAPX1属于植物过氧化物酶超家族成员,与其他物种过氧化物酶体APX相似性较高,C端具1个跨膜区。荧光定量PCR结果表明,BnAPX1在苎麻的根、茎中段、茎尖、茎皮、幼叶各部位均有表达,其中幼叶表达量最高,且该基因BnAPX1受重金属镉诱导上调表达,可能在重金属镉胁迫防御中起重要作用。
The full-length cDNA of APX gene was cloned from ramie(Boehmeria nivea L.)through RT-PCR and RACE methods according to the APX gene sequence of ramie transcriptome, named as BnAPX1. The full length cDNA of BnAPX1 was 1 201 bp, which has an open read frame of 870 bp, encoding a protein of 289 amino acids. Bioinformatic analysis showed that BnAPX1 protein shares the characters of plant peroxidase family. Amino acids sequence analysis suggested that BnAPX1 protein is highly similar to other APX proteins and is possessed of a transmembrance domain at its C-terminal. Real-time PCR analysis showed that BnAPX1 mRNA is expressed in the organs in ramie root, middle stem, shoot tip, stem bark, and young leaf. And the expression of BnAPX1 was the highest in youn leaf. BnAPX1 was also found to be up-regulated by Cd, which suggested that it might play a role on Cd stress.
朱守晶,周精华,揭雨成,等.苎麻抗坏血酸过氧化物酶基因的克隆和表达分析[J].植物遗传资源学报,2013,14(5):879-884.
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