Cloning and expression of human of MAdCAM-1 in E. coli as well as purification of recombinant protein

The extracellular fragment of cDNA encoding human MAdCAN-1 was amplified from plasmid pUC21/hMAdCAM-1 and then cloned into vector pQE30 in fusion form by PCR and recombinant DNA techniques. The recombinant vector pQE30-MAd was transformed into E. coli M15. After IPTG induction, a fusion protein with relative molecule mass (M_(r)) of 29 000 was expressed, representing 50% of total bacterial protein in E. coli. SDS-PAGE assay showed the products has high purity after purified by Ni-NTA affinity chromatography column. The recombinant hMAdCAM-1 was successfully obtained, this might provide an experimental basis for the function research and mAb preparation hereafter.