以含有IPT基因的中间载体pSG516为基础,采用酶切的方法获得IPT-Nos核酸片段,以水稻品种9311为材料,采用PCR的方法克隆出种子中特异表达的醇溶谷蛋白RP-6基因启动子,并将此启动子连接到pCAMBIA1300上,构建pCAMBIA1300-pRP-6载体,将IPT-Nos核酸片段插入到pCAMBIA1300-pRP-6,构建了pCAMBIA1300-pRP-6-IPT-Nos双元表达载体.In order to obtain transgenic rice which express exogenous IPT gene in developing seed and to further investigate the function of cytokinin in rice seed development,we constructed the binary vector pCAMBIA1300-pRP-6-IPT-Nos.Firstly IPT-Nos fragment was obtained from pSG516 by cutting PSG516 plasmid with Nco I and Spe I restriction enzymes.Secondly,developing endosperm RP-6 gene specifically expressed in developing endosperm was selected and it′s promoter(pRP-6) was isolated from rice genomic DNA(cultivar 9311) by using PCR amplification.Thirdly pRP6 was inserted into pCAMBIA1300,with pCAMBIA1300-pRP-6 obtained.Finally,plant expression vector pCAMBIA130-pRP-6-IPT-Nos was constructed after inserting IPT-Nos fragment into pCAMBIA1300-pRP-6.福建省青年科技人才创新项目(2005J004)资
