摘要： 为研究细胞分裂素在水稻籽粒发育中的作用,构建了p1300- pPG-5α-IPT-Nos 植物表达载体,并对水稻进行遗传转化。以水稻品种9311为材料,采用PCR法获得种子中特异表达的 PG-5α 基因启动子,并将此启动子与p1300相连接,构建p1300- pPG-5α 载体,用 Nco Ⅰ和 Spe Ⅰ双酶切pSG516,获得 IPT-Nos 核酸片段,然后将此核酸片段插入到p1300- pPG-5α 中,构建p1300- pPG-5α-IPT-Nos 植物表达载体。以水稻品种日本晴愈伤组织为材料,采用农杆菌介导的方法进行遗传转化。成功构建了植物表达载体p1300- pPG-5α-IPT-Nos,获得了阳性率为81.3%转 IPT 基因群体。获得转基因株系后,将为进一步筛选高效表达株系及观察其籽粒生长发育过程提供基础。

关键词: PG-5α 启动子, IPT, p1300- pPG-5α-IPT-Nos, 水稻, 遗传转化

Abstract: In order to specifically express IPT gene in the endosperm of developing rice seed, the binary vector p1300- pPG-5α-IPT-Nos was constructed, firstly, the promoter of PG-5α was obtained by PCR amplification from rice genomic DNA(variety:9311)and p1300- pPG-5α was constructed by inserting pPG-5α into p1300, then the IPT-Nos fragment was obtained by cutting pSG516 with Nco Ⅰ and Spe Ⅰ.After IPT-Nos inserted into p1300- pPG-5α, p1300- pPG-5α-IPT-Nos was constructed. pPG-5α-IPT-Nos was transformated into Oryza sativa (subsp.japonicacv.Nipponbare) cells by agrobacterium mediated method and regenerative seedlings were obtained.The results of PCR showed that about 81.3% seedlings were positive.

Key words: The promoter of PG-5α, IPT, p1300- pPG-5α-IPT-Nos, Oryza sativa, Transformation

中图分类号: 

• Q78