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为利用RNA介导的病毒抗性策略,培育抗性稳定或抗多烟草蚀纹病毒(Tobacco etch virus,TEV)株系的转基因植株,采用RT-PCR及5’-RACE方法克隆了烟草蚀纹病毒山东分离物TEV-SD1的全基因组序列。TEV-SD1全基因组核苷酸序列长度为9494 bp,包含1个9165 bp的开放阅读框架(open reading frame,ORF),编码3054个氨基酸。将TEV-SD1基因组序列与GenBank中已公布的4个TEV全基因组序列和11个外壳蛋白(coat protein,CP)基因序列比对分析发现,各分离物CP基因间的核苷酸和氨基酸序列平均相似性分别为96.65%和98.31%,高于其它功能基因间的相似性;各分离物CP基因3’端核苷酸序列相似性平均为96.55%,高于5’端序列。聚类分析发现TEV在自然界中的分子变异与其寄主关系密切。
In order to utilize RNA-mediated virus resistance strategies to develop transgenic plants resistant or resistant to Tobacco etch virus (TEV) strains, tobacco etchings were cloned by RT-PCR and 5’-RACE Whole genome sequence of virus Shandong isolate TEV-SD1. The full-length nucleotide sequence of TEV-SD1 is 9494 bp in length and contains a 9165 bp open reading frame (ORF) encoding 3054 amino acids. The alignment of TEV-SD1 genomic sequences with published 4 TEV genome sequences and 11 coat protein (CP) gene sequences in GenBank revealed that the nucleotide and amino acid sequences of CP genes of each isolate averaged The similarities were 96.65% and 98.31%, respectively, which were higher than those of other functional genes. The 3 ’nucleotide similarity of CP gene in each isolate was 96.55%, higher than that of the 5’ end. Cluster analysis revealed that the molecular variation of TEV in nature is closely related to its host.