|
|
|
| Prokaryotic expression of N-terminal antigenic domain of duck plague virus gB |
| Hua-Qi Pan Nan Wang Lei Liu Jiang-Chun Hu |
| |
|
|
|
|
Abstract According to the antigenic analysis of duck plague virus (DPV) gB protein , one pair of primers were designed, with which the gene fragment coding the high antigenic domain of DPV N-terminal gB protein was amplified from the DPV genome. The segment was cloned into pET32a vector to obtain the recombinant pET-gB1 plasmid. The recombinant plasmid was transformed into E. coli BL21, and expressed in very high level as inclusion body after induced with IPTG. The expressived product analyzed by SDS- PAGE and Western blotting. The result indicated that the fusion protein (pET-gB1) has specific antigenicity. The inclusion body was solubilized in urea (8mol/L) . The purified protein was obtained by use of His•Bind affinity chromatography. Mice were inoculated transcutaneous three times at a two-weeks interval with fusion protein and normal saline. To evaluate the prophylaxtic efficacy of fusion protein , Titers of antibodies was detected by ELISA and proliferation of lymphocytes were determined by MTT. The results indicated that fusion protein could elicit specific humoral immune and cellular immune responses.
|
|
Received: 05 December 2007
|
|
Corresponding Authors:
Jiang-Chun Hu
|
|
|
|
|
|
|
