Title: Expression of huainan porcine interleukin-2 in Escherichia coli and bioactivity analysis.
Abstract: [Objective] This study developed prokaryotic expression system which can express porcine interleukin-2(pIL2)in Escherichia coli,and identified the bioactivity of the recombinant protein.[Method] Porcine IL-2 gene was amplified with recombination plasmid pGEM-pIL2 as a template,porcine IL-2 was cloned into the prokaryotic expression vector of pGEX-6p-1 and transformed into E.coli BL21(DE3)competent cells,which can express fused protein GST-pIL2 in E.coli BL21(DE3)by IPTG inducing.[Result] A 420 bp fragment was amplified by PCR.DNA sequencing result showed that the sequence of the recombinant prokaryotic expression plasmid pGEX-pIL2 in the reading frame and the ligation part was correct.The expressed product was identified by SDS-PAGE and Western-blot.The expressed protein was about 42 ku and it could react with rabbit against porcine IL-2 monoclonal antibody.MTT essay confirmed that the expressed product can enhance lymphocyte proliferation.[Conclusion] Porcine IL-2 gene can express in E.coli,and the expressed protein has bioactivity.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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