大豆GmPR10基因启动子的克隆及序列分析
首发时间:2015-12-11
摘要:GmPR10基因是病程相关蛋白PR10(Pathogenesis-related Proteins 10)在大豆中的同源基因。为探明大豆GmPR10基因的表达调控规律,应用PCR技术从大豆抗疫霉根腐病品种"绥农10号"中克隆了GmPR10基因上游2235bp的启动子序列pGmPR10,定向替换pBI121载体的CaMV35S组成型启动子,构建植物表达载体pBI121/pGmPR10/GUS,并转化烟草叶盘。GUS染色结果表明,pGmPR10具有启动功能,且受聚乙二醇(PEG)、低温(4℃)、水杨酸(SA)、茉莉酸(JA)和脱落酸(ABA)诱导表达,因此推测GmPR10基因可能参与植物激素调节植物生长发育的过程,以及生物胁迫和非生物胁迫条件下植物对环境响应的过程。此外,利用PLACE和PlantCARE在线启动子预测工具分析pGmPR10,结果表明:pGmPR10含有启动子的一般结构TATA-box和CAAT-box,光应答元件,生长素和细胞分裂素响应元件,热激元件,低温应答元件,干旱应答元件以及ABA、SA、JA应答元件等。上述研究为GmPR10基因的功能解析奠定了理论基础。
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Cloning and Sequence Analysis of the GmPR10 Gene Promoter from Soybean (Glycine max L.)
Abstract:GmPR10 was homology with the PR10 (pathogenesis-related proteins 10) gene in other plants. In order to investigate the expression and regulation of the GmPR10 gene in soybean, the promoter sequence which contained a 5′-flanking upstream 2235bp sequence of GmPR10 gene was isolated from the genomic DNA of soybean cultivars "Suinong 10"(with high resistance against Phytophthora sojae in Heilongjiang, China) by PCR method, defined as pGmPR10. It was directionally replaced CaMV35S constitutive promoter of the vector pBI121. The plant expression vector of pBI121/pGmPR10/GUS was then constructed by linking the cloned fragment to GUS gene, used for transformation of tobacco. Then the expression of downstream reporter GUS gene was drived. GUS staining results showed that, pGmPR10 owing to start function, and the expression of GmPR10 gene may be induced by drought (PEG), cold, salicylic acid (SA), jasmonic acid (JA) and abscisic acid (ABA). Therefore, it was speculated that GmPR10 gene may be involved in the process of plant growth and development with the regulation of plant hormones, as well as the process under abiotic and abiotic stresses responses to the environment.In addition, sequence analysis by PLACE and PlantCARE revealed that pGmPR10 contained promoter general structure TATA-box and CAAT-box, light response element, auxin and cytokinin response element, heat shock element, cold response element, drought responsive element and ABA, SA, JA response elements.
Keywords: soybean GmPR10 promoter cloning GUS staining
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