紫花苜蓿MsNAP启动子克隆及转基因分析  

Cloning of MsNAP Promoter from Medicago sativa and Transgenic Analysis

作　　者：穆致远 谢利娟[2] 郭涛[1] 袁建波[1] 李殷睿智 晁跃辉[1] Mu Zhiyuan;Xie Lijuan;Guo Tao;Yuan Jianbo;Li Yinruizhi;Chao Yuehui(Turfgrass Research Institute,College of Forestry,Beijing Forestry University,Beijing,100083;School of Applied Chemistry and Biotechnology,Shenzhen Polytechnic,Shenzhen,518055)

机构地区：[1]北京林业大学林学院草坪研究所,北京100083 [2]深圳职业技术学院应用化学与生物技术学院,深圳518055

出　　处：《分子植物育种》2018年第20期6688-6694,共7页Molecular Plant Breeding

基　　金：北京林业大学科技创新计划项目(2017ZY16);深圳市科技计划项目(JCYJ20160331151245672)共同资助

摘　　要：启动子位于基因上游,起着调控基因转录的作用,是研究基因表达特征和功能的重要元件。为了研究MsNAP启动子活性,探究MsNAP基因功能,进而研究高产优质的紫花苜蓿,造福畜牧业,本试验利用染色体步移法,成功获得了紫花苜蓿MsNAP基因翻译起始位点上游1 390 bp序列,通过生物信息学分析,发现该区域存在一个完整的启动子,含有多种顺式作用元件。利用DNA重组技术,成功构建了MsNAP启动子表达载体,通过农杆菌介导技术成功转入到烟草中。选取衰老与未衰老的烟草叶片,进行葡萄糖苷酸酶(glucuronidase, GUS)染色分析,结果显示,在衰老叶片中GUS活性远远高于未衰老叶片,表明MsNAP启动子在衰老的叶片中具有较高的转录调控活性,从而为深入研究MsNAP启动子活性和分析MsNAP基因的功能提供了基础。Promoter is a fragment upstream of a gene and an important element in analyzing expression pattern and gene function, playing a role in regulating transcription. This research was aimed to investigate the activity MsNAP promoter, the function ofMsNAP gene and study Medicago sativa with high yield and good quality, thus to benefit animal husbandry. In this study, a 1 390 bp upstream fragment of MsNAP translation starting site was isolated by genome walking method. It was found by bioinformation analysis that there was a completed promoter containing many cis-elements in this region. The expression vector of MsNAP promoter was successfully con- structed by DNA recombination technology and transferred into tobacco plants by Agrobacterium tumefacien-me- diated transformation. The glucuronidase （GUS） activity was analyzed in senescent and non-senescent leaves of one tobacco plant. The results showed that the activity of GUS in senescent was much higher than that in non-senes- cent leaves, which indicated that MsNAP promoter had a stronger activity of transcriptional regulation in senescent leaves, Therefore, it would provide a basis for further study of MsNAP promoter activity and analysis of the function of MsNA P gene.

关 键 词：MsNAP 启动子 紫花苜蓿 衰老 转基因 

分 类 号：Q943.2[生物学—植物学] S541.9[农业科学—作物学]