海州香薷(Elsholtzia haichowensis Sun)细胞壁转化酶基因启动子(EhcwINVP)的克隆及活性分析  

Isolation and Activity Analysis of Cell Wall Invertase Gene Promoter( Ehcw INVP) from Elsholtzia haichowensis Sun

作　　者：赵静[1] 蔡深文[1] 徐仲瑞[1] 熊治廷[1] 

机构地区：[1]武汉大学资源与环境科学学院,武汉430079

出　　处：《植物科学学报》2016年第3期420-429,共10页Plant Science Journal

基　　金：国家自然科学基金项目(21477093;31270432);生物质资源化学与环境生物技术湖北省重点实验室资助项目(HBRCEBL2013-2014004)~~

摘　　要：以海州香薷基因组DNA为模板,通过hiTAIL-PCR和walking技术扩增得到其细胞壁转化酶基因启动子(Ehcw INVP)片段,长度为1727 bp。生物信息学分析结果表明,该启动子片段中含有多个对脱落酸、赤霉素、细胞分裂素等激素以及对干旱、低温、重金属铜等逆境胁迫响应相关的顺式作用元件。将通过克隆得到的Ehcw INVP序列替换p CAMBIA1301载体上驱动GUS报告基因表达的Ca MV35S启动子序列,构建Ehcw INVP融合GUS的植物表达载体Ehcw INVP::GUS。转基因拟南芥植株的组织化学分析结果表明,海州香薷细胞壁转化酶基因启动子序列具有驱动GUS基因表达的功能,且在10μmol/L铜胁迫下,转基因拟南芥植株叶和根中的GUS活性分别约是对照组的1.7倍和1.5倍。Cell wall invertase gene promoter,Ehcw INVP（ 1727 bp）,was amplified from the genomic DNA of Elsholtzia haichowensis Sun by hi TAIL-PCR and genome walking PCR.Bioinformatics analysis showed that the promoter fragment contained multiple cis-acting elements involved in the regulation of hormones including abscisic acid,gibberellin,cytokinin,and tolerance to drought, low temperature, and heavy metals such as copper. The p CAMBIA1301 binary vector was digested with restriction enzymes to remove the 35 S promoter that drove GUS expression and replaced with Ehcw INVP sequences to generate the construct Eh Ncw INVP： ： GUS. Histochemical analyses of transgenic Arabidopsis thaliana L.plants indicated that the Ehcw INVP promoter was capable of driving GUS expression,and GUS activity in the leaves and roots of transgenic A. thaliana plants increased by about 1.7 and1.5 times that of the control under copper stress,respectively.

关 键 词：海州香薷 细胞壁转化酶 启动子 顺式作用元件 

分 类 号：Q78[生物学—分子生物学]