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应用RT-PCR技术克隆了水稻瘤矮病毒(RGDV)中国广东信宜分离物(RGDV-C)的基因组S9片段,测定了全序列并进行了生物信息学分析。结果表明,RGDV-C S9片段全长共有1202bp(登录号AY556483),含有一个长的开放阅读框,这一开放阅读框编码一个由323个氨基酸残基组成的多肽,推测分子量约35.6kDa,与泰国分离物(RGDV-T)的全序列相比,它们的核苷酸长度相等,核苷酸同源性为98.1%,氨基酸同源性为98.5%。RGDV S9片段编码的Pns9蛋白在植物呼肠孤病毒属内未发现同源蛋白,其功能尚待确定。利用NCBI的BLAST查找与比较,发现Pns9与伯氏疏螺旋体(Borrelia burgdorferi)ATP依赖的Clp蛋白水解酶组分[ATP-dependent Clp prote-ase proteolytic component(clpP-1)]有21.8%的氨基酸序列同源性。
The genomic S9 fragment of RGDV-RGDV-C was cloned by RT-PCR. The full-length sequence was sequenced and bioinformatics analysis was performed. The results showed that the full-length RGDV-C S9 fragment was 1202bp in length (accession number AY556483) and contained a long open reading frame. This open reading frame encoded a polypeptide consisting of 323 amino acid residues with a predicted molecular mass of about 35.6 kDa. Compared to the full sequence of the Thai isolate (RGDV-T), their nucleotide lengths were equal, with nucleotide homology of 98.1% and amino acid identity of 98.5%. The Pns9 protein encoded by the RGDV S9 fragment has no homologous protein found in the plant reovirus and its function remains to be determined. Using BLAST search and comparison of NCBI, it was found that Pns9 has 21.8% amino acid sequence with ATP-dependent Clp prote-ase proteolytic component (clpP-1) of Borrelia burgdorferi Homology.