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以已知植物抗病基因的保守区域为基础设计简并引物,从冬瓜抗病材料B227基因组中分离NBS-LRR类抗病基因同源序列,共获得32条特异序列,其中17条具有完整开放阅读框。这17条序列的核苷酸同源性变化范围为56.4%~99.8%,氨基酸的同源性变化范围为46.2%~100.0%。序列相似性分析结果表明,这些冬瓜抗病基因同源序列均包含P-loop,Kinase-2和GLPL等保守结构域。与已知抗病基因构建系统进化树,将这17个序列分为2个亚组,均属于TIR-NBS-LRR类抗病基因。本研究将为抗病基因全长序列克隆、功能分析及定位等研究提供帮助。
Degenerate primers were designed based on the conserved regions of known plant disease resistance genes. NBS-LRR homologous sequences were isolated from B227 genome of winter melon, and 32 specific sequences were obtained, of which 17 were completely open Reading frame. The nucleotide sequences of these 17 sequences varied from 56.4% to 99.8%, and amino acid homologies varied from 46.2% to 100.0%. Sequence similarity analysis showed that the homologous sequences of these resistance genes contained conserved domains such as P-loop, Kinase-2 and GLPL. The phylogenetic tree was constructed based on the known resistance genes. The 17 sequences were divided into two subgroups and belong to the TIR-NBS-LRR resistance gene. This study will help clone the full-length sequence of disease resistance genes, analyze their functions and locate them.