Phylogenetic analysis and prokaryotic expression of NMAAP1 derived from BCG-activated murine macrophages

Abstract

BCG-activated macrophages exerted anti-tumor activities. Cell surface molecules play an important role in mediating endocytosis by macrophages. In the previous study, we identified a group of 454 membrane proteins specifically expressed on BCG-activated mouse macrophages, including a protein named NMAAP1 (novel macrophage activated associated protein). In this study, we aligned the full-length nucleotide sequences of NMAAP1 and its homologous sequences to construct its phylogenetic tree, and cloned the NMAAP1 cDNA from BCG-activated macrophages to generated NMAAP1 fusion protein in Escherichia coli. Purified the fusion protein were applied for generation of polyclonal antibodies. Western-blotting detection showed that the polyclonal antibodies have high specificities to recognize target protein.