Cloning, Expression, Purification And Identification Of The Interaction Molecular Of Foxp3-C22orf37

The fork head transcription factor FoxP3 is specificly expressed in T cells witll regulatory.and assoeiated with immunity suppression of Treg.CD4~+CD25~+ Treg cells and transcription factor FOXP3 have been the focus of research in the field of immunoregulation mechanisms in the last ten years.It has been confirmed that CD4+CD25+Treg cells play a important role in immunoregulation and FOXP3 has emerged as a crificM regulator of the development and function of CD4~+CD25~+Treg cells.Recently,it is consider ed that FOXP3 is a more specific marker to distinguish CD4~+CD25~+Treg cells from CD4~+CD25~-T cells at least in mouse.There are many factors to influence the induction and expression of FOXP3 and many kinds of molecules are involved in immane regulation by FOXP3,so it is helpful to provide more insight into the role of FOXP3 in immunoregulation.we have successfully constructed engineering bacteria DH5αincluding pQE30-C22orf37 gene and induced to express intereste protein C22orf37 with 1mM IPTG.The fusion protein shows good reactivity to anti-his antibody. Interested protein expressed in the form of inclusion body.Inclusion body after washing was dissolved in 8M urea and was dialysed to binding buffer.The purity of purified interested protein C22orf37 by Ni affinity chromatography exceed to 90%.High titer of immunoglobulin could not be induced when was C22orf37 used to immune mice.The interested genes were cloned into pQE30 vector and expressed in Escherichia coli.Several interested proteins were purified by affinity chromatography.The high titer immunoglobulin could be induced when several recombinant proteins were used to immune BALB/c mice or New Zealand rabbits.The titer of antiseras all reach to 1:10~4 and the one coming from the group immunized with recombinant protein C22orf37 exceeded to 1:10~4. Practical immunocytochemistry in some unhomologization cell line indicated that this moleculor has exprexssed in some unhomologization cells.Successfully constructed and purified C22orf37 protein of human mature C22orf37.we immune the mouse to pruoduce high antibody.Moreover,we find that this possiblely expressed on multiple human tumor cells.