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Virologica Sinica, 21 (4) : 385-389, 2006
Research Article
Cloning and Expression of VP1 Gene of Swine Vesicular Disease Virus
1. Faculty of Bioengineering and Chemical Engineering, Kunming University of Science and Technology, Kunming 650224, China
2. Technology Center of Yunnan Entry-Exit Inspection and Quarantine Bureau, Kunming 650228, China
3. Xishuangbanna Veterinary Station, Jinghong 666100, China
 Correspondence:
(453.16KB)  
Abstract
The VP1 gene of Swine vesicular disease virus (SVDV) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) yielding a product of 849bp cDNA fragment. Using T-A cloning technique, the PCR product was cloned into pMD18-T vector. The purified VP1 gene was subcloned into the pBAD/Thio TOPO vector and the plasmid was identified by PCR. It was sequenced to confirm the authenticity of the sequence and orientations. SDS-PAGE and Western blotting revealed that the VP1 protein was expressed in Escherichia coli LGM194 at a high level and the recombinant fusion protein contained a N-terminal HP-Thioredoxin and a C-terminal polyhistidine tag. The optimal amount of the expressed fusion protein was 16% of total bacterial protein after being induced with L-arabinose at 0.002%concentration for 5 hours. It had a molecular mass of approximately 47.13 kDa and was immunologically reactive. The recombinant protein will be characterized and tested in an enzyme-linked immunosorbent assay (ELISA) format for potential application in the serodiagnosis of swine vesicular disease.
  Published online: 20 Jul 2006
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