作物学报 ›› 2015, Vol. 41 ›› Issue (04): 524-530.doi: 10.3724/SP.J.1006.2015.00524
王红梅1,张昌泉1,李钱峰1,辛世文2,刘巧泉1,*,徐明良1,3
WANG Hong-Mei1,ZHANG Chang-Quan1,LI Qian-Feng1, SUN Samuel Sing-Min2,LIU Qiao-Quan1,*,XU Ming-Liang1,3
摘要:
提高外源蛋白在特定目标组织器官中的表达量是转基因植物研究与开发的核心技术之一。谷蛋白是水稻种子中最主要的贮藏蛋白, 其表达具有严格的时空特异性。为进一步研究谷蛋白信号肽序列在指导基因表达中的作用,本研究克隆了水稻谷蛋白GluA-2 基因的启动子及其信号肽编码序列, 并与GUS 报告基因编码区融合, 构建了分别含有和不含有信号肽的表达载体p13GG 和p13GSG; 经农杆菌介导法分别转入同一水稻品种中, 获得了20 多个独立转化子, PCR 证明外源基因都已整合进了水稻基因组中。Northern 杂交结果表明, 融合GluA-2 信号肽编码序列可显著提高GUS 基因在水稻胚乳中的转录; 利用GUS 特异的抗体进行Western 杂交分析, 显示该信号肽序列可显著提高外源蛋白在转基因水稻胚乳中的积累, 但是其所指导表达的GUS 蛋白在水稻胚乳中并没有表现出相应的活性, 其机制有待进一步深入解析。相关结果对于水稻品质改良基因工程研究以及以水稻种子作为生物反应器高效表达外源蛋白具有重要的指导作用。
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