Basic Information
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Targeting strategy
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Gene targeting strategy for B-hADAM9 mice.
The exons 2-19 of mouse Adam9 gene that encode the extracellular domain were replaced by human ADAM9 exons 2-19 in B-hADAM9 mice. -
Protein expression analysis
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Western blot analysis of ADAM9 protein expression in homozygous B-hADAM9 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM9 mice (H/H), and then analyzed by western blot with species-specific anti-ADAM9 antibody. 40 μg total proteins were loaded for western blotting analysis. ADAM9 protein was detected in the heart, liver, spleen, lung, kidney, brain and thymus of wild-type C57BL/6 mice and homozygous B-hADAM9 mice, as the antibody is cross-recognize both human and mouse ADAM9.
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mRNA expression analysis
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Strain specific analysis of ADAM9 mRNA expression in wild-type C57BL/6 mice and B-hADAM9 mice by RT-PCR. Brain and lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM9 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human ADAM9 primers. Mouse Adam9 mRNA was detectable only in wild-type C57BL/6 mice. Human ADAM9 mRNA was detectable only in homozygous B-hADAM9 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 Adam9 mRNA was detectable in wild-type C57BL/6 mice. Human ADAM9 mRNA was detectable in homozygous B-hADAM9 mice.
Protein expression analysis:
ADAM9 protein was detected in the heart, liver, spleen, lung, kidney, brain and thymus of wild-type C57BL/6 mice and homozygous B-hADAM9 mice, as the antibody is cross-recognize both human and mouse ADAM9.
