稻属NBS-LRR类基因的克隆及抗稻瘟病基因的表达

Cloning of NBS Gene from Oryzae and Expression of Rice Blast Resistance Gene

利用已克隆植物的R基因NBS序列中保守基序设计简并引物进行PCR扩增是克隆NBS有效的方法。从广东普通野生稻HLW028中克隆出3条序列,经同源性分析均为NBS,序列号为DQ272573～DQ272575。从NCBI中下载普通野生水稻和功能已知的水稻NBS-LRR类基因,与本试验所提交的NBS进行聚类分析。这些NBS可分为5大类,其中DQ272574是一个新的NBS基因类型。从野生水稻中克隆NBS基因存在P-loop、RNBS-A、kin-2、RNBS-B、RNBS-C和PLAL的基序。RT-PCR结果表明,培矮64中的Pikh的表达量比2种野生稻中高。从培矮64中扩增出1个完整读码框的cDNA。

It was an effective method to clone NBS using the degenerate primers designed by the conserved motifs of NBS of the cloned R genes. Three sequences were cloned from wild rice HLW028 and identified as NBS. The accession numbers were DQ272573--DQ272575. The NBS of wild rice and the functionally known NBS of rice were downloaded. Phylogenetic analyses were conducted with the three submitted NBS and other downloaded NBS. The NBS could be divided into five groups, DQ272574 was a new kind of NBS. The submitted NBS contained the motifs such as P-loop, RNBS-A, kin-2, RNBS-B, RNBS-C and PLAL. RT-PCR indicated that the expression of Pikh in Peiai64 was higher than that in 2 wild rice. A cDNA with intact open reading frame was amplified from Peiai64.