DNA sequence and prokaryotic expression analysis of vitellogenin from Antheraea pernyi
In this study, the DNA sequence of vitellogenin from Antheraea pernyi (Ap-Vg) was identified and its functional domain (30-740 aa, Ap-Vg-1) was expressed in Escherichia coli BL21 (DE3) cells. The recombinant Ap-Vg-1 proteins were purified and used for antibody preparation. The results showed that the intact DNA sequence of Ap-Vg consisted of 8124 bp including 6 exons (2258, 205, 982, 879, 184 and 829 bp) and 5 introns (84, 78, 410, 1055 and 795 bp) and was highly homologous to the vitellogenin from Antheraea yamamai. SDS-PAGE and western blot analysis demonstrated that a 85 KD recombinant protein was successfully expressed in E. coli cells and its expression was not remarkably changed under induction by different IPTG concentration. The titre of antibodies raised against rabbits was about 1:7800 which was determined by ELISA.