目的 通过lncRNA测序鉴定三叶青Tetrastigma hemsleyanum烟草花叶病毒蛋白基因。方法 利用大肠杆菌重组技术表达三叶青烟草花叶病毒蛋白，并采用生物信息学方法进行序列分析。结果 三叶青烟草花叶病毒蛋白基因包括复制酶、RNA聚合酶、运动蛋白、带电蛋白和外壳蛋白等基因；三叶青烟草花叶病毒RNA聚合酶、运动蛋白（MP）、带电蛋白和外壳蛋白（CP）可实现大肠杆菌重组表达，表明三叶青烟草花叶病毒的lncRNA测序结果准确；三叶青烟草花叶病毒CP、MP、RNA聚合酶和带电蛋白基因cDNA总长度分别为480、807、1 425、123 bp，三叶青烟草花叶病毒CP、MP、RNA聚合酶和带电蛋白分别由159、268、474、40个氨基酸组成；三叶青烟草花叶病毒CP、MP、RNA聚合酶为亲水性蛋白质，带电蛋白为疏水性蛋白质；三叶青烟草花叶病毒CP、MP、RNA聚合酶二级结构由α螺旋、β-片层、无规则卷曲构成，带电蛋白二级结构由β-片层和无规则卷曲构成，三级结构均为单体，分别定位于叶绿体、叶绿体、细胞质和线粒体中。三叶青烟草花叶病毒CP、MP、RNA聚合酶和带电蛋白与烟草花叶病毒亲缘关系较近，同源性达98%以上。结论 三叶青烟草花叶病毒具有烟草花叶病毒的结构特征，氨基酸序列及核酸序列与同源物种相似度高，在进化上高度保守。

Objective To identify the protein gene of tobaccos mosaic virus in Tetrastigma hemsleyanum by sequencing of lncRNA. Methods Proteins was expressed by Escherichia coli recombinant technology, and the sequence was analyzed by bioinformatics methods. Results The results showed that the protein genes of tobaccos mosaic virus in T. hemsleyanum included replicase, RNA polymerase, motor protein, charged protein and coat protein genes; The recombinant expression of its RNA polymerase, movement protein (MP), charged protein and coat protein (CP) could be realized in E. coli, which indicated that the sequencing result of lncRNA was accurate; The total cDNA length of CP, MP, RNA polymerase and charged protein genes of tobaccos mosaic virus in T. hemsleyanum were 480, 807, 1 425 and 123 bp, respectively. The CP, MP, RNA polymerase and charged protein were composed of 159, 268, 474 and 40 amino acids, respectively; The CP, MP, RNA polymerase was hydrophilic protein, and the charged protein was hydrophobic protein; The secondary structure of CP, MP, RNA polymerase was composed of alphahelix, extendedstran and randomcoil and the secondary structure of charged protein was composed of extendedstran and randomcoil. Their tertiary structures were all monomer, which were located in chloroplast, chloroplast, cytoplasm and mitochondria respectively. The CP, MP, RNA polymerase and charged protein of T. hemsleyanum were closely related to tobacco mosaic virus, with more than 98% homology. Conclusion TMV of T. hemsleyanum had the structural characteristics of tobacco mosaic virus, whose amino acid sequence and nucleic acid sequence were highly similar to those of homologous species and highly conservative in evolution.

R286.12

国家自然科学基金资助项目(31860084);国家自然科学基金资助项目(31960079);国家自然科学基金资助项目(32060092);江西省现代农业产业技术体系建设专项(JXARS-13-赣东站)