摘要
为研究植物Lea (lateembryogenesisabundant)蛋白基因启动子在种子中的特异性表达 ,通过PCR扩增 ,从棉花(Gossypiumhirsutumcv .Coker312 )中克隆了Lea蛋白基因家族中D - 113基因上游 10 2 4bp的调控序列。DNA序列分析结果表明 ,该片段与已报道的Lea蛋白基因同一家族该基因的对应序列同源性达 90 %以上。将该启动子序列与GUS基因融合 ,构建成表达载体后 ,通过基因枪轰击导入到经ABA诱导处理的棉花胚性愈伤组织和油菜种子以及棉花的根、茎、叶中 ,组织化学分析结果表明 ,D - 113基因启动子在胚中特异性表达。
To study the expression of late embryogenesis abundant gene in seeds, the 1 024bp 5′flanking sequence of D -113 gene, a late embryogenesis abundant gene of Gossypium hirsutum cv. Coker 312, was cloned by PCR. The similarity compared with the sequence of Lea protein gene family published was 92.50%. There are three putative ABREs and one enhancer like which riches A/T in the promoter. The promoter was fused to the β glucuronidase gene to form pLDⅡ. Via a particle bombardment, pLDⅡ was introduced into embryogenic calli of cotton and seeds of Brassica napus which were all treated with abscisic acid for 3d before bombardment, also into roots, stems and leafs of cotton. Transient expression was measured histochemically as spot number 24h after bombardment. GUS sexpression was observed in the seeds of Brassica napus and the embryogenic calli of cotton, but not found in roots and leaves of cotton. Those results indicated that the expression of D -113 gene promoter was embryo specific.
基金
国家转基因植物研究与产业化专项课题资助 (资助号 :ZJY -OA -0 1)
