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Updated: Jun 13, 2026
Phage Phenomics: Physiological Approaches to Characterize Novel Viral Proteins
Published on: June 11, 2015
Phylogenetic analysis and prokaryotic expression of NMAAP1 derived from BCG-activated murine macrophages
Xiangfeng Zhao1, Linbing Zhang, Dongmei Yan
1Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun 130021, China. zxf184@163.com
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.
Insights
Researchers identified novel macrophage activated associated protein 1 (NMAAP1) on BCG-activated macrophages. Antibodies were generated against NMAAP1, confirming its specific detection in macrophage research.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Macrophages activated by Bacillus Calmette-Guérin (BCG) exhibit anti-tumor properties.
- Cell surface molecules are crucial for macrophage-mediated endocytosis.
- A previous study identified 454 membrane proteins on BCG-activated mouse macrophages, including NMAAP1.
Purpose of the Study:
- To characterize the novel macrophage activated associated protein 1 (NMAAP1).
- To generate specific antibodies for NMAAP1 detection.
Main Methods:
- Phylogenetic analysis of NMAAP1 nucleotide sequences.
- Cloning of NMAAP1 cDNA from BCG-activated macrophages.
- Expression and purification of NMAAP1 fusion protein in Escherichia coli.
- Generation of polyclonal antibodies against NMAAP1 fusion protein.
- Western-blotting to assess antibody specificity.
Main Results:
- The phylogenetic tree for NMAAP1 and its homologs was constructed.
- NMAAP1 cDNA was successfully cloned and a fusion protein was generated.
- Polyclonal antibodies against NMAAP1 were successfully produced.
- Western-blotting confirmed the high specificity of the generated antibodies for NMAAP1.
Conclusions:
- NMAAP1 is a novel protein expressed on BCG-activated macrophages.
- Specific polyclonal antibodies against NMAAP1 have been developed.
- These antibodies are valuable tools for further investigating NMAAP1's role in macrophage function.
