Function of GMFB and GMFG in brain development

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(1) Production of Specific Antibodies against Glia Maturation Factor beta (GMFB) and gamma (GMFG).Recombinant proteins of rat GMFB and GMFG were expressed by E. coli (BL21(DE3)) using pAED4 expression vector, and purified by column chromatography. Polyclonal antibodies were obtained by immunization to New Zealand White Rabbits. Specific antibodies were isolated by affinity chromatography. By western blot, no cross-reactivity was detected between GMFB antibody and GMFG, or between GMFG antibody and GMFB.(2) Establishment of Enzyme Immunoassay System for GMFB and GMFG.Sandwich EIA system was established by using the specific antibodies against GMFB and GMFG respectively. These EIAs enabled us to identify immunoreactive GMFB and GMFG in human plasma and rat organs. GMFB was expressed in brain, spleen, thymus, lung and intestine, and GMFG was expressed in thymus and spleen. These results correspond to the results of western blot and immunohistochemical analyses.(3) Cloning of genomic DNA of human and mouse GMFG.Genomic DNA of human and mouse GMFG were cloned. Sequence analyses revealed that both human and mouse GMFG gene were consisted by 7 exons.