Cloning and protein expression of mouse MAdCAM-1 spliced variant
Obtaining the spliced mMAdCAM-1 cDNA from mesenteric lymph nodes of BALB/c mice by RT-PCR, cloning the cDNA into the procaryotic expressive vector pGEX-2T and expressing the fusion protein. After identified by DNA sequencing, the fusion protein was expressed and mainly existed as inclusion bodies. After purification, we prepared the multi clonal-antibody to the fusion protein. The spliced mMAdCAM-1 protein was identified by the multi clonal-antibody and standard monoclonal antibody against mMAdCAM-1 with Western blot.