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Abstract According to the gene sequence of African swine fever virus (ASFV ) VP73 in the gene bank , VP73 was synthetized and cloned into pMD18-T vector. The VP73 gene of African Swine Fever Virus (ASFV) was amplified from DNA by PCR, the product of which was a 1188 bp DNA segment. The purified VP73 gene was subcloned into pBAD/Thio TOPO vector. The recombinant plasmid was identified by PCR. It was sequenced to confirm the correct sequences and the correct junctional orientations of the inserted VP73 gene. The recombinant protein will be purified by proBond TM protein. The results of SDS-PAGE revealed that the VP73 protein was expressed in pBAD/Thio TOPO vector in a high level .The optimal amount of the expressed fusion protein is 30% of total bacterial protein after being induced with L-arabinose at 0.002%concentration for 4 hours. It had a molecular mass of approximately 60 kDa and immunologically reactive activity. The VP73 protein was expressed in pBAD/Thio TOPO vector in a high level . The recombinant protein purified by proBond TM protein and tested in an enzymelinked immunosorbent assay (ELISA), it results that the VP73 fusion protein has good reactinogenicity, which establishes the fundament of using the recombinant protein to prepare ASFV serological diagnostic reagent and vaccines.
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Received: 23 June 2006
Published: 25 November 2006
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