摘要
通过特异PCR扩增技术,以拟南芥基因组DNA为模板,克隆了rd29A基因上游1 600 bp的胁迫诱导型启动子序列。将此1 600 bp的调控序列与GUS基因连接构建植物表达载体pBI101-rd29A-GUS,并用农杆菌介导法转基因拟南芥。定量PCR结果显示,干旱胁迫下转基因纯合株系中rd29A显著上调表达。GUS组织化学染色及GUS定量分析结果表明,在ABA、甘露醇和NaCl等胁迫处理下,GUS活性增强,尤其是ABA胁迫处理。说明rd29A启动子可以增强逆境下GUS基因的表达,可作为一种诱导型启动子应用于提高作物抗逆性的基因工程研究中。
About 1600bp stress inducible promoter of rd29A from Arabidopsis thaliana genomic DNA was cloned by PCR. The plant expression vector pBI101-rd29A-GUS was constructed with this regulatory region linked up with GUS gene,and then transferred to Arabidopsis by Agrobacterium tumefaciens system. The expression level of rd29A was up-regulated under drought stress in homozygous seedlings. Histochemical analysis and quantitative analysis results showed that the GUS activity was enhanced under ABA、NaCl and mannitol treatments. Therefore, rd29A prompter can strengthen the expression of GUS gene under stresses, and can be used as an inducible promoter in gene engineering for stress resistance improvement of crops.
作者
柳娜
杨文雄
王世红
张雪婷
杨长刚
LIU Na;YANG Wenxiong;WANG Shihong;ZHANG Xueting;YANG Changgang(Institute of Wheat,Gansu Academy of Agricultural Sciences,Lanzhou Gansu 730070,China)
出处
《甘肃农业科技》
2019年第5期40-46,共7页
Gansu Agricultural Science and Technology
基金
国家自然科学基金项目(31560390)
甘肃省科技重大专项(17ZD2NA016-7)
甘肃省小麦产业技术体系(GARS-01-01)
公益性行业农业科研专项(201503125-1)资助
