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从云南省弥勒县采集典型病株接种蔓陀萝，植斑分离，经免疫电镜（ＩＥＭ）鉴定为ＴＭＶ（Ｔｏｂａｃｃｏｍｏｓａｉｃｖｉｒｕｓ），在红花大金元品种中以ＴＭＶ普通株作对照进行致病力测定，确定为强毒株系。以烟草花叶病毒ＲＮＡ云南强毒株系为模板，根据国内外研究结果自行设计、合成寡核苷酸引物，通过ＲＴ—ＰＣＲ体外扩增，Ｅ．ｃｏｌｉ菌株ＤＨ５α克隆，获得了云南烟草５４ＫＤ（ＴＭＶ复制酶）全基因片段。顺序分析表明：该基因含１４２５个核苷酸，编码４７５个氨基酸，与国外发表的Ｕ１株系比较，ＤＮＡ同源率为９９．０％，氨基酸同源率为９８．９％，与国内普通株比较，ＤＮＡ同源率为９９．１％，氨基酸同源率为９８．９％。
The typical strains were collected from Maitreya County in Yunnan Province and inoculated with M. difficiles and the plaques were isolated and identified as TMV (Tobacco mosaic virus) by immunoelectron microscopy (IEM). The virulence of TMV strains , Identified as virulent strains. The virulent strains of tobacco mosaic virus Yunnan virulent strain were used as a template, and oligonucleotide primers were designed and synthesized according to the research results both at home and abroad. The amplified products were amplified by RT-PCR in vitro. coli strain DH5α cloned, obtained Yunnan tobacco 54KD (TMV replicase) whole gene fragment. Sequence analysis showed that the gene contained 1425 nucleotides and encoded 475 amino acids. Compared with the published U1 strains, the DNA homology was 99.0% and the amino acid homology was 98.9% The DNA homology was 99.1% and the amino acid homology was 98.9%.