大豆花叶病毒外壳蛋白基因的克隆和其在大肠杆菌中的表达

Cloning and Expression of Coat Protein Gene of Soybean Mosaic Virus in Escherichia coli

通过多聚酶连锁反应(PCR),我们合成了包括病毒外壳蛋白基因在内的病毒基因组3′端区域,并完成了其全部序列分析,比较SMV(北京分离物)和SMV-N株的序列发现:外壳蛋白基因的核苷酸序列同源性达93.4%,其氨基酸序列同源性高达98.5%,就基因组3′端非编码序列而言,其同源性达88.8%。Western blot分析结果表明所克隆的cDNA片段在大肠杆菌JM107中能表达正常的病毒外壳蛋白。

The 3'-terminal genomic region of the Beijing isolate of Soybean Mosaic Virus (SMV-BJ) has been cloned through technique of polymerase chain reaction (PCR). We have analysized the nucleotide sequence of 3' region of SMV-BJ genome. Comparisons of the nucleotide and deduced amino acid sequences of SMV-BJ coat protein gene with those of SMV-N strain show 93.4% and 98.5% identity between them, respectively. Alignments of the 3' non-coding sequence in pair with that of SMV-N strain show homology of 88.8%. It has been found that the SMV coat protein gene is expressed in E. coli by western blot analysis. The coat protein produced in E. coli has the same electrophoretic mobility as SMV coat protein.