Ohno T, Aoyagi M, Yamanashi Y, Saito H, Ikawa S, Meshi T, Okada Y
J Biochem. 1984 Dec;96(6):1915-23. doi: 10.1093/oxfordjournals.jbchem.a135026.
The sequence of about 4,500 nucleotides of the internal part of tobacco mosaic virus (TMV)-tomato strain (L) RNA has been newly determined using cloned cDNAs. Together with the previously determined partial sequences at both ends, the entire sequence of the 6,384 nucleotide genome has been completed. The 130K (1,115 amino acids), 180K (1,615 amino acids), 30K (263 amino acids) and coat protein (158 amino acids) cistrons are located at residues 72-3442, 72-4922, 4906-5700, and 5703-6182 on the genome, respectively. Sequence polymorphism was not observed except for heterogeneity in the length of the A cluster near the 3' end. The homology of the nucleotide sequences of TMV-L and TMV-vulgare, a common strain, is about 80% on average. Remarkable differences between them were found in a part of the N-terminal portion of the 130K/180K protein and the C-terminal portion of the 30K protein. A new method for cDNA cloning was developed by which the cDNA of the 5'-terminus of viral RNA can be cloned efficiently.
利用克隆的互补DNA(cDNA),新测定了烟草花叶病毒(TMV)番茄株系(L)RNA内部约4500个核苷酸的序列。结合之前测定的两端部分序列,完成了6384个核苷酸基因组的全序列测定。130K(1115个氨基酸)、180K(1615个氨基酸)、30K(263个氨基酸)和外壳蛋白(158个氨基酸)的顺反子分别位于基因组上的第72 - 3442位、72 - 4922位、4906 - 5700位和5703 - 6182位残基处。除了3'端附近A簇长度的异质性外,未观察到序列多态性。TMV-L与常见株系TMV-普通株的核苷酸序列同源性平均约为80%。在它们之间,130K/180K蛋白的N端部分和30K蛋白的C端部分存在显著差异。开发了一种新的cDNA克隆方法,通过该方法可以高效克隆病毒RNA 5'端的cDNA。