论文部分内容阅读
根据烟草花叶病毒(Tobacomosaicvirus,TMV)U1株系(TMV-U1)序列,人工合成引物,用RT法合成cDNA后,通过PCR技术扩增并克隆了烟草花叶病毒蚕豆株系(TMV-B)的外壳蛋白(CP)基因和3'端非编码区。DNA序列测定结果表明,外壳蛋白基因全长480个碱基,编码158个氨基酸,3'端非编码区全长204个碱基,与TMV-U1株系的同源率为100%。将CP基因及3'端非编码区插入pGEMEX-1的T7基因10中,转入E.coli后诱导表达,聚丙烯酰胺凝胶电泳分析呈现一特异的蛋白带,Western免疫检测证明,该特异带与TMV抗血清呈阳性反应。
According to the sequence of Tobacco mosaic virus (TMV) U1 strain (TMV-U1), primers were synthesized and amplified by RT-PCR. The TMV-B ) Coat protein (CP) gene and 3 ’non-coding region. The results of DNA sequence analysis showed that the coat protein gene was 480 bases in length and encoded 158 amino acids. The 3 ’non-coding region was 204 bases in length. The homology was 100% with TMV-U1 strain. The CP gene and the 3 ’noncoding region were inserted into the T7 gene of pGEMEX-1 10 and transferred into E. coli. After induced by E. coli, polyacrylamide gel electrophoresis showed a specific protein band. Western immunization showed that the specific band was positive with TMV antiserum.