Construction and expression of recombinant hMAdCAM-1/GST fusion gene expression vector

ObjectiveTo construct and express a recombinant expression vector bearing hMAdCAM-1/GST fusion gene. MethodshMAdCAM-1 cDNA 5'-end 615 bp fragment encoding hMAdCAM-1 Ig-like domains was amplified by PCR from plasmid pUC21/hMAdCAM-1 and then inserted into pGEM-T plasmid. Determined by auto-sequencing, the target gene fragment was cloned into prokaryotic expression vector pGEX-2T in fusion form and transformed into E.coli DH5a. ResultsSDS-PAGE analysis showed that a new protein band with molecule weight of 52 000 u appeared as the expected size, representing 31% of the total bacterial protein. Western-blot indicate that the expressed fusion protein could bind with antiserum against hMAdCAM-1 specifically. ConclusionThe successful construction and expression of recombinant vector pGEX-2T-MAd might provide material for the further research about hMAdCAM-1.