Cloning and expression of a fragment of swine HEV ORF2 AG02 in E.coli and the Development of Indirect ELISA with

China Biotechnology

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Cloning and expression of a fragment of swine HEV ORF2 AG02 in E.coli and the Development of Indirect ELISA with

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Abstract Nest RT-PCR was used to amplified swine hepatitis E virus(SHEV) ORF2-AG02 fragment.AG02 was inserted into pET32a vector and the recombinant expression plasmid PET-HEV was constructed. PET-HEV was transferred into BL21(DE3) and induced by 1mM IPTG, the SDS-PAGE result showed that the 33kDa recombinant protein was expressed. The Western blot analysis showed that the recombinant protein has a good antigenicity with positive SHEV serum. An indirected ELISA was established with the purified recombinant protein, after the square matrix titrate determination best density is 2.43μg/mL, the blood serum best dilution ratio is 1:40,Compares with ten reagents boxes, proved we establish the ELISA method has the high specificity and the sensitivity.

Key words： ORF2 Protoplast fusion insect cell ELISA HEV

Received: 24 March 2006 Published: 25 August 2006

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. Cloning and expression of a fragment of swine HEV ORF2 AG02 in E.coli and the Development of Indirect ELISA with. China Biotechnology, 2006, 26(08): 37-41.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I08/37

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