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The eukaryotic expression and identification of Fcγ-Der f2 fusion protein and examination of its biological activity

LIN Li-hui,CUI Yu-bao,KONG Cun-quan,et al. 1.Department of Clinical Laboratory,the First People' s Hospital of Shanghai,Shanghai 200080,China; 2.Yancheng Vocational & Technical Health College,Jiangsu 224006 China  
Objective To construct the human Fcγ-Der f 2 chimeric gene,and to study the expression and effect of Fcγ-Der f 2 fusion protein.Methods Amplified The cDNA encoding Der f2 from pMD19-T-Der f2 vector was amplified and the flexible linker sequence was set at the 5′-end.Fcγand linker-Der f2 was inserted into pDisplay expression vector.The new pDisplay-Fcγ-Der f2 vector was transfected into HEK293T cells by Attractene reagent.And the expression of fusion Fcγ-Der f 2 protein was determined by immunofluorescence and western blot.Results The results of the sequence and restriction enzyme analysis showed that pDisplay -Fcγ-Der f2vector was successfully constructed.The immunofluorescence demonstrated that the recombinant fusion protein was correctly expressed in HEK293T cells.At the same time,the Western blot showed the antigenicity of Der f2 in the fusion protein and the molecular weight of this protein was 40×10~3.Conclusion The Fcγ-Der f 2 fusion gene was constructed successfully and the fusion protein is of biological activity.
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