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利用RT -PCR技术 ,合成并扩增了水稻条叶枯病毒 (RStV)中国云南分离物基因组组分 3的全长cDNA。将PCR产物克隆在载体 pCRⅡ上 ,进行全序列测定。将所得核苷酸序列及其所推导的氨基酸序列与日本分离物T进行同源性比较 ,结果表明 ,在核苷酸水平上 ,两分离物的 5′端非编码区序列相同 ,vORF、vcORF及基因间非编码区序列的同源性分别为 97 6 %、96 8%及 87 6 % ,而 3′端非编码区同源性为 98 9% ,仅有一个核苷酸不同 ;在氨基酸水平上 ,vORF及vcORF编码蛋白的同源性分别为 99 1%和 98 5 %。可见编码区的大小及其氨基酸序列和两末端序列都是很保守的。序列分析显示 ,两个RStV分离物的RNA3均有双义编码特性。因此 ,中国云南分离物Y与日本分离物T可能有很近的亲缘关系。
Using RT-PCR technique, the full-length cDNA of genomic component 3 of Yunnan isolate was synthesized and amplified from rice leaf blight virus (RStV). The PCR product was cloned into the vector pCRII and sequenced. The results of nucleotide homology comparison between the obtained nucleotide sequence and deduced amino acid sequence and Japanese isolate T showed that the 5 ’non-coding region sequences of the two isolates were the same at the nucleotide level, and the vORF, vcORF And the homology of the non-coding region of the gene were 97.6%, 96.8% and 87.6% respectively, while the homology of the 3 ’non-coding region was 98.9% with only one nucleotide difference. In the amino acid At the same level, the homologies of vORF and vcORF encoded proteins were 99 1% and 98 5%, respectively. It can be seen that the size of the coding region and its amino acid sequence and both terminal sequences are very conservative. Sequence analysis showed that RNA3 of both RStV isolates had a double-sense coding property. Therefore, there may be a close genetic relationship between Yunnan isolate Y and Japanese isolate T.