Basic Information
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Description
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- MCEMP1 (mast cell-expressed membrane protein 1) is a 22 kDa mast cell specific protein expressed in lung.
- Human MCEMP1 is 187 amino acids (aa) in length. It is a type II transmembrane protein that contains an 85 aa N-terminal cytoplasmic region and an 81 aa C-terminal extracellular domain (aa 107‑187). There is one potential splice variant that shows a 22 aa substitution for the C-terminal five amino acids. Over aa 106‑187, human MCEMP1 shares only 52% aa identity with the presumed mouse MCEMP1.
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Targeting strategy
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Gene targeting strategy for B-hMCEMP1 mice.
The exons 4-7 of mouse Mcemp1 gene that encode signal peptide, extracellular domain and transmembrane domain is replaced by human counterparts in B-hMCEMP1 mice. The 3’UTR region of the mouse gene are replaced by human counterparts. The chimeric MCEMP1 expression is driven by endogenous mouse Mcemp1 promoter, while mouse Mcemp1 gene transcription and translation will be disrupted.
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mRNA expression analysis
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Strain specific analysis of MCEMP1 mRNA expression in wild-type C57BL/6 mice and B-hMCEMP1 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hMCEMP1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human MCEMP1 primers. Mouse Mcemp1 mRNA was detectable in wild-type C57BL/6. Human MCEMP1 mRNA was detectable only in homozygous B-hMCEMP1 mice but not in wild-type mice.
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Protein expression analysis
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Western blot analysis of MCEMP1 protein expression in homozygous B-hMCEMP1 mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hMCEMP1 mice (H/H), and then analyzed by western blot with anti-MCEMP1 antibody (abcam, ab121447). 40 μg total proteins were loaded for western blotting analysis. MCEMP1 was detected in lung, heart, spleen and kidney in wild-type mice and homozygous B-hMCEMP1 mice, as the antibody is cross-recognized between human and mouse MCEMP1.
