拟南芥rd29A启动子克隆及其在甘蔗抗逆转基因中应用研究初探.pdfVIP

拟南芥rd29A 启动子的克隆及其在甘蔗抗逆转基因中的应用初探1 张积森、廖辉煌、林晓坤、陈如凯、张木清2 （福建农林大学农业部甘蔗生理生态与遗传改良重点实验室，福州350002 ） 要：从 南芥(Arabidopsis thaliana)基因组DNA 克隆了长度为420bp 的rd29A 基因启动子, 序列分析表明该启动子具有两个干旱、高盐和低温响应顺式作用元件(DRE),其它主要 控区 域也存在。用rd29A 启动子构建了具有绿色荧光蛋白(GFP)基因的植物表达载体，通过基因 枪导入法转化甘蔗愈伤组织，在转化后3d 和5d，NaCl 胁迫条件下，用荧光显微镜观察GFP 基因在愈伤组织表达水平.结果表明 rd29A 启动子是NaCl 诱导型启动子,可满足在盐胁迫条 件下使目的基因在转基因甘蔗中进行表达的目的。 关键词：甘蔗；rd29A ；gfp ；抗逆 Cloning of rd29A Gene Promoter from Arabidopsis thaliana and Its Preliminary Application in the Sugarcane Transformation of Stress resistance Zhang J.-X., Liao H.-H., Lin X.-K., Chen R.-K. and Zhang M.-Q. (Key Lab of Eco-Physiology Genetic Improvement for Sugarcane under Ministry of Agriculture, Fujian Agri. Forestry Univ., Fuzhou, 250002) Abstract :The 420 bp region of rd29A gene promoter from Arabidopsis thaliana genome was amplified by PCR technique. Sequence analysis showed that the fragment contained two cis-acting elements including dehydration responsive elements (DRE) and ABA responsive elements (ABRE) and the main regulation motifs. The plant expression vector pPrd-S65T was constructed with this fragment linked up with green fluorescent protein (GFP) gene controlled by rd29A promoter, which was transferred to sugarcane callus by bombardment microproj ectiles. The expressions of gfp in the transformed sugarcane callus treated with 250 mmol/L NaCl for3 days and 5 days, were detected by fluorescent microscope.The result indicated that the cloned rd29A promoter could be applied in the transgenic sugarcane for the stress resistance. Key words : sugarcane; rd29A gene promoter ;green fluorescent protein (GFP) gene ; Stress resistance 1基金资助：国家“863”计划课题糖料新品种选育课题 （200 1AA24 1191 ）；国家自然科学基金 ） 以及教育部高等学校博士学科点专项科研基金项目 （20040389009 ） 2通讯作者(Author for correspondence): 张木清( 1966-)、男，福建周宁人，汉族、博士、博士生导师、 教授，主要研究方向：作物生