玉米盐诱导型启动子克隆及其植物表达载体构建.docVIP

Botanical Research 植物学研究, 2012, 1, 30-35 /10.12677/br.2012.12005 Published Online July 2012 (/journal/br) Isolation of Salt-Inducible Promoter in Maize and Construction of Plant Expression Vector* Yajie Zheng, Di Feng, Yi Wei, Jinliang Liu, Baolei Jia, Yanhua Zhang, Hongyu Pan, Shihong Zhang # College of Plant Sciences, Jilin University, Changchun # Email: {yajiebaobei, fengdi0804}@163.com, zhang_sh@ Received: May 7th, 2012; revised: May 28th, 2012; accepted: Jun. 6th, 2012 Abstract: The upstream nucleotide sequence of maize salt-inducible protein kinase, named STY, was isolated from the genomic DNA of B73 by PCR. Promoter sequence analysis by PlantCARE showed that it had some typical cis-elements, including TATA box and CAAT box. In addition, there were two salt responsive elements GT1-motif, various phytohormone responsive elements and stress responsive elements in the promoter sequence. The fusion gene of STY promoter and GUS were constructed and named pCAM-STY. The vector was trans- formed into Agrobacterium EHA105. The vector containing STY promoter was transferred into tobacco by Agrobacterium tumefaciens mediated method, function of STY promoter was analyzed by identifying activation of GUS using histochemistry staining. The results indicated that the promoter activated expression of GUS re- port gene, it laid the foundation to resolve the serious multi-stress problems of agriculture in northeast of China. Keywords: Maize; Salt-Inducible; Promoter; Sequence Analysis; Transformation; Functional Verification 玉米盐诱导型启动子克隆及其植物表达载体构建* 郑亚杰，冯 頔，魏毅，刘金亮，贾保磊，张艳华，潘洪玉，张世宏# 吉林大学植物科学学院，长春 Email: {yajiebaobei, fengdi0804}@163.com, zhang_sh@ # 收稿日期：2012年 5月 7日；修回日期：2012年 5月 28日；录用日期：2012年 6月 6日 摘 要：首先提取 B73玉米(Zea mays)的基因组 DNA，以此为模板利用 PCR的方法克隆玉米盐诱导蛋 白激酶(STY)基因上游 1675 bp，将其命名为 STY。从网站 PlantCARE上的启动子预测工具在线对克隆 到的启动子进行分析，结果表明：STY启动子中含有多种调控元件，典型的调控元件有：TATA-box， CAAT-box等等。另外还有 2个盐诱导顺式作用元件 GT1-motif和一些冷、干旱胁迫诱导有关的顺式作 用元件，以及功能未知的元件。然后将克隆到