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High Soluble Expression and Antigenicity of VP1 Gene of Foot-and-mouth disease virus |
XIE Qing-mei;LI Shao-li;CHEN Li;QIN Jian-ping;MA Jing-yun;BI Ying-zuo; CAO Yong-chang |
College of Animal Science, South China Agricultural University, Guangzhou 510640, China |
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Abstract Foot-and-mouth disease virus (FMDV) VP1 gene was amplified by RT-PCR. The VP1 fragment was inserted into plasmid pMBX to obtain recombinant plasmid pMBX-VP1. The pMBX-VP1 was transformed into Escherichia coli BL-21(DE3) to induce VP1 gene fusion expression with 1 mmol/L Isopropylthio-β-D-galactoside (IPTG). The fusion protein band with molecular weight of 58 kD was visible on the SDS-PAGE gel. The density scanning showed that the largest amount of the fusion protein was 35.5% of total bacterial protein. The amounts of the soluble form of the fusion in supernatant of lysate was up to 31.2%, and 6.8% in sediment. Western blot result shawed that the expression products could specifically reacted with the antisera against FMDV. The soluble fusion protein was purified by 50%Ni-NTA affinity chromatography and used as an antigen, and FMDV antibody could be detected by ELISA method.
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Received: 13 September 2005
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Corresponding Authors:
CAO Yong-chang
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