Basic Information
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Targeting strategy
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Gene targeting strategy for B-hMMP9 mice.
The exons 1-13 of mouse Mmp9 gene that encode the full-length protein were replaced by human MMP9 exons 1-13 in B-hMMP9 mice. -
Protein expression analysis
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Western blot analysis of MMP9 protein expression in homozygous B-hMMP9 mice. Various tissue lysates were collected from C57BL/6N wild-type (+/+) mice and homozygous B-hMMP9 (H/H) mice, and then analyzed by western blot with anti-MMP9 antibody. 40 μg total proteins were loaded for western blotting analysis. MMP9 was detected in bone marrow, lymph node, lung and spleen both from wild-type and homozygous mice, as the antibody is cross-recognized both for human mouse MMP9. The molecular weight of mouse MMP9 is slight bigger than that of human MMP9, which might be mainly due to the number of amino acids and the post-transcriptional modification.
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mRNA expression analysis
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Strain specific analysis of MMP9 mRNA expression in wild-type C57BL/6N mice and B-hMMP9 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hMMP9 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human MMP9 primers. Mouse Mmp9 mRNA was detectable only in wild-type C57BL/6N mice. Human MMP9 mRNA was detectable in homozygous B-hMMP9 mice but not in wild-type mice. And the PCR products were confirmed with Sanger sequencing.
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Summary
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mRNA expression analysis:
Mouse Mmp9 mRNA was detectable in lung of wild-type C57BL/6N mice. Human MMP9 mRNA was detectable in lung of B-hMMP9 mice and was confirmed with Sanger sequencing.Protein expression analysis:
MMP9 was both detectable in bone marrow, lymph node and spleen from WT and homozygous B-hMMP9 mice, as the antibody is cross-recognized both for human and mouse MMP9.
