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首页> 外文期刊>African Journal of Biotechnology >Molecular cloning and expression analysis of an Mn-superoxide dismutase gene in sugarcane
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Molecular cloning and expression analysis of an Mn-superoxide dismutase gene in sugarcane

机译：甘蔗中Mn超氧化物歧化酶基因的分子克隆和表达分析

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Superoxide dismutases (SODs) play an important role in stress-tolerance in plants. In this study, for the first time, a full-length cDNA sequence of?MnSOD?gene, termed asSc-MnSOD?(GenBank accession number: GQ246460), was obtained in sugarcane. Sequence analysis revealed that?Sc-MnSOD?gene was 919 bp long, including a 702 bp ORF, the 5’ UTR of 99 bp and 3’UTR of 118 bp. It encoded the 233 amino acid residues with a molecular weight of 25.3 KD and isoelectric point of 7.11. Protein domain prediction indicated that besides the conserved domain in MnSOD, Sc-MnSOD?also had the signal peptides at the sites of 1 to 26 aa at the N-terminus. With SubLoc v1.0, Sc-MnSOD was localized in the mitochondria. In homology analysis, theMnSOD?genes from different plant species were rather conservative. SDS-PAGE analysis and enzyme activity assay showed that the prokaryotic expression product was a fusion protein with SOD enzyme activity of 726 U·mg-1. Real-time qPCR analysis demonstrated that the?expression of?Sc-MnSOD?gene?was greatly induced by the?Ustilgao scitaminea, firstly induced and then inhibited by H2O2, and slightly influenced by SA, which suggested that it may play a role in the clearing of active oxygen and thus in disease resistance mechanism in sugarcane.
机译：超氧化物歧化酶（SOD）在植物的耐逆性中起重要作用。在这项研究中，首次在甘蔗中获得了称为MnSODα基因的全长cDNA序列，称为Sc-MnSODα（GenBank登录号：GQ246460）。序列分析表明，Sc-MnSOD基因长919 bp，其中ORF为702 bp，5'UTR为99 bp，3'UTR为118 bp。它编码了233个氨基酸残基，分子量为25.3 KD，等电点为7.11。蛋白质结构域的预测表明，除了MnSOD中的保守结构域外，Sc-MnSODα在N端的1至26aa位点也具有信号肽。使用SubLoc v1.0，Sc-MnSOD定位于线粒体中。在同源性分析中，来自不同植物物种的MnSOD 2基因相当保守。 SDS-PAGE分析和酶活性分析表明，原核表达产物是一种融合蛋白，SOD酶活性为726 U·mg-1。实时qPCR分析表明，Ustilgao scitaminea极大地诱导了“ Sc-MnSOD”基因的表达，首先被H2O2诱导然后被其抑制，然后被SA轻微地影响，这表明它可能在以下方面起作用。清除活性氧，从而影响甘蔗的抗病机理。

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