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通过逆转录PCR(RT-PCR)及快速扩增cDNA末端(RACE)技术从中国白梨品种雪花梨中克隆到S16-RNase基因的全长cDNA序列(GenBank接受号为DQ991388).该cDNA克隆总长1101bp,包含1个完整的开放阅读框,编码228个氨基酸.S16-RNase表现出梨S-RNase基因的基本结构特征,即其具有5个保守区(C1,C2,C3,RC4及C5)和1个高变区.在推导氨基酸水平上,S16-RNase与梨其他S-RNase基因的相似性为63%至74%,但与苹果S9-RNase的相似性高达95%.通过多序列比对构建进化树,分析梨S16-RNase与蔷薇科植物其他S-RNase基因的遗传进化关系.结果表明,S16-RNase与苹果亚科S-RNase基因形成一个亚科特异而非种特异的S-RNase基因类群;且不同S-RNase基因间存在属内种间遗传距离远于属间种间遗传距离现象.
The full-length cDNA sequence of S16-RNase gene (GenBank accession number is DQ991388) was cloned from Chinese white pear snowberry by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) 1101bp, contains a complete open reading frame encoding 228 amino acids.S16-RNase shows the basic structural characteristics of pear S-RNase gene, which has 5 conserved regions (C1, C2, C3, RC4 and C5) and 1 hypervariable region.The similarity of S16-RNase and other pear S-RNase genes was 63% to 74% at the deduced amino acid level, but 95% similarity with apple S9-RNase.Through multiple sequence alignment The phylogenetic relationship was established between S16-RNase and other S-RNase genes in Rosaceae plants.The results showed that S-RNase and S-RNase genes of apple subfamily formed a subfamily specific but not species-specific S-RNase Gene groups; and there is an intraspecific genetic distance between different S-RNase genes far away from the genetic distance between species.