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Abstract To construct prokaryotic expression vector containing CVN-LP1 gene ，and identify the expression of CVN-LP1 protein using Western blot assay。 Methods： pET-CVN and pET-LP1 was restriction endonuclease digested by EcoRⅠ and HindⅢ, and the CVN gene was inserted into pET-LP1 plasmid 。The vector was transformed into BL21(DE3) to construct a prokaryotic expression system 。The expressed product was identified by SDS-PAGE and Western blot assay。 Results： The recombinant plasmid pET-CVN-LP1 was constructed， restriction endonuclease digestion and PCR identification proved that the CVN was correctly cloned into vector pET-LP1 。SDS-PAGE and Western-blot analysis showed that CVN-LP1 was successfully expressed in Ecoli BL21(DE3) ，The relative molecular mass (Mr) of the expression protein was 20kD，according with the predicted Mr value。And the expression yield was about 16.86% of total bacterial protein。 Conclusion： The successful expression of CVN-LP1 gene in Ecoli provides a support for further study of the anti-HIV agent。
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Received: 09 October 2006
Published: 25 March 2007
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