Construction and identification of mutant recombinant plasmids of human suppressor of cytokine signaling

Objective To construct the mutant recombinant plasmids express the CDS and 3 ' UTR gene of human suppressor of cytokine signaling (SOCS)3.Methods pEGFP-C1-SOCS3 wild type plasmid as the template,the CDS and 3' UTR gene of SOCS3 was obtained by PCR amplification.According to the SOCS3 mRNA and miR-155 prediction of binding sites,respectively,to design SOCS3 gene mutation types:3' UTR-1,3' UTR-2 and CDS.By two rounds PCR will get the mutations of the SOCS3 fusion fragments which were cloned into pEGFP-C1 expression vector.The fusion recombinant plasmids were confirmed by PCR and DNA sequen-cing.Results The mutant recombinant plasmids express the CDS and 3 ' UTR gene of human SOCS3 wereconstructed and identified by DNA sequencing as same as the prediction.Conclusions The successfully con-structed pEGFP-C1-SOCS3 mutant plasmids are useful to study the interaction of SOCS3 and miR-155. Key words: SOCS3; General recombination; PCR; Mutation