| Based on the TGB1 sequence of Cymbidium mosaic virus (CyMV, GenBank Accession No: HQ681906.1),a pair of specific primers was designed for amplifying the TGB1 gene from the total RNA of Datura stramonium leaves infected with CyMV. DNA sequencing showed that the TGB1 gene length was 702 bp and encoded 233 amino acid residues. The TGB1 gene was subcloned into a prokaryotic expression vector pET32a (+). By inducing with 1.0 mmol/L IPTG at 25℃, the recombinant TGB1 gene was expressed in E. coli BL21(DE3) and its product was identified as a specific band of 44 ku by SDS-PAGE. Polyclonal antibodies against recombinant TGB1 protein were obtained by hypodermal injection of rabbits. The titre of antiserum was 1:25 600. |
