2004•Zhongguo yufang shouyi xuebaoRequires access
Expression of DNA encoding VP2 of MDPV in a pichia pastoris and checking of the expressed products
Lou Hua
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Abstract
Two recombinant plasmids pMD18-T-M-Q VP2 and pMD18-T-M-26 VP2 were digested with KpnI and SacII . The VP2 inserts were sub-cloned into the expression vector pPICZαA, and recombinant expression plasmids pPICZαA-M-Q VP2 and pPICZαA-M-26 VP2 were obtained.The two recombinant expression plasmids were used to transform into the Pichia Pastoris X-33 and the inserted target genes were expressed.The expressed products were analyzed and checked by SDS-PAGE and Western-blot. Western-blot analysis showed one band of approximately equal intensity that corresponded to the expressed proteins of approximately 73 kD, conforming to theoretical estimate. In addition, an unexpected result was found in that. Western-blot analysis showed two bands;One band was the expected VP2 band, the other was approximatey 60 kD