摘要
利用已知的水稻条纹叶枯病毒的序列设计了12对引物,利用这些引物经RT-PCR获得了5条序列,经BLAST分析,它们均与水稻基因组内的序列完全不匹配,可以用来作为干扰序列。将这5个序列构建入由pUC19改造的一段内含子两侧各含有一对同尾酶的干扰载体中,然后再将整个RNA干扰片段构建入经改造的以玉米泛素Ubi为启动子、以生物安全性的bar基因为选择标记的植物双元表达载体pCAMBIA1300中,经农杆菌介导对黄淮稻区主栽水稻品种圣稻13进行遗传转化,为抗条纹叶枯病水稻新品种的培育提供了候选材料。
Twelve pairs of primers were designed according to the known sequences of rice stripe virus,and 5 sequences were obtained by RT-PCR.By BLAST analysis,no similar sequences in rice genome were found matching these five sequences,so they could be used as the interference sequences.A modified pUC19 vector inserted a fragment as intron with a pair of isocaudarners on each side was used as the interference vector.After inserting the interference sequences into the interference vector separately,the complete interference fragments were integrated into the modified binary vector pCAMBIA1300 using ubiquitin as promoter and bar gene as selectable marker.The transgenic rice seedlings were obtained by Agrobacterium-mediated transformation to rice variety Shengdao 13.This provided candidate materials for breeding new rice varieties resistant to rice stripe virus.
出处
《山东农业科学》
2011年第2期1-6,共6页
Shandong Agricultural Sciences
基金
国家转基因植物新品种培育重大专项(编号:2009ZX08001-010B)资助
