农杆菌介导的转高赖氨酸蛋白基因(sb401)水稻T4代分析  

Analysis on T_4 Progeny of Transgenic Rice with Lysine-Rich Protein Gene (sb401) Mediated by Agrobacterium tumefaciens Transformation

作　　者：李科[1,2] 王世全[1,2] 吴发强[1,2] 李双成[1,2] 邓其明[1,2] 王玲霞[1,2] 梁越洋[1,2] 李平[1,2] 

机构地区：[1]四川农业大学水稻研究所,四川成都611130 [2]四川农业大学西南作物基因资源与遗传改良教育部重点实验室,四川雅安625014

出　　处：《中国水稻科学》2008年第2期131-136,共6页Chinese Journal of Rice Science

基　　金：国家863计划资助项目(2006AA10Z1D1);教育部长江学者和创新团队发展计划资助项目(IRT0453)

摘　　要：通过农杆菌介导的方法用富赖氨酸蛋白基因（sb401）转化粳稻品种日本晴，获得了独立的10个转化株系，对转化株系进行连续自交，通过筛选得到9个纯合的T4代转化株系。Southern blotting分析发现，整合位点是随机的，并为低拷贝（1～3个）。TAIL-PCR扩增得到8个T—DNA侧翼序列，并定位于日本晴的7条染色体上。蛋白质和氨基酸测定分析发现，sb401基因对各株系的蛋白质、赖氨酸和其他氨基酸组分的提高起到了一定的作用。将杂交结果与T-DNA插入位置结合分析发现，在低拷贝的情况下，表达量的差异不明显。The lysine-rich protein gene sb401 had been successfully introduced into a japonica rice variety Nipponbare by Agrobacterium-mediated transformation. Out of ten independent transformed lines, nine homozygous T4 generation transgenic lines were obtained through consecutive selling. Southern blotting analysis of T4 lines showed that the integration sites of the foreign gene were random with low copy numbers （1 to 3）. TAIL：PCR analysis showed that eight T-DNA flanking sequences were located on seven chromosomes. Detection of protein and amino acids contents of T4 lines showed that sb401 gene contributed, to some extent, to the increase of lysine and other amino acids contents in transgenic lines. The analysis combining blotting and T-DNA position showed that the expression difference was not obvious when there were low copies in transgenic rice lines.

关 键 词：转基因水稻 位置效应 富赖氨酸蛋白基因 热不对称交错聚合酶链式反应 赖氨酸含量 蛋白质含量 

分 类 号：S511[农业科学—作物学]