Expression ofσC gene of muscovy duck reovirus,MW9710 strain,by using pMAL-p2X system
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Graphical Abstract
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Abstract
To obtain recombinant Muscovy duck reovirus(MDRV) _σC protein by prokaryotic expression for further study on its functions,the encoding sequence of the _σC gene from the MDRV MW9710 strain was amplified with RT-PCR and inserted into pMAL-p2X vector to establish the prokaryotic expression plasmid.After transforming the plasmid,pMAL-p2X-_σC,into E.coil BL21(DE3),the bacteria were induced by IPTG.The protein was expressed efficiently in both soluble and insoluble forms,and was positively recognized by the reference serum.The SDS-PAGE and western-blot analysis showed the recombinant pMAL-p2X-_σC produced had an apparent molecular weight of 72KDa.The soluble _σC protein was purified and obtained using MBP-affinity chromatography.The result provided a foundation for the gene’s epitope identification and development of a clinical diagnostic kit in the future.
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