Basic Information
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Targeting strategy
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Gene targeting strategy for B-hADA mice.
Exons 1-12 of the mouse Ada gene, which encodes the full-length protein, were replaced by human ADA exons 1-12 in B-hADA mice. -
mRNA expression analysis
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Strain specific analysis of ADA mRNA expression in wild-type C57BL/6 mice and B-hADA mice by RT-PCR. Spleen RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hADA mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human ADA primers. Mouse Ada mRNA was detectable in wild-type C57BL/6 mice. Human ADA mRNA was detectable in homozygous B-hADA mice but not in wild-type mice.
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Protein expression analysis
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Strain specific ADA expression analysis in homozygous B-hADA mice by western blot. Tissues were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADA mice (H/H), and analyzed by western blot with anti-ADA antibody. ADA was detectable in homozygous B-hADA mice.
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Summary
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mRNA expression analysis:
Mouse Ada mRNA was detectable in the spleen of wild-type C57BL/6 mice. Human ADA mRNA was detectable in B-hADA mice.Protein expression analysis:
ADA was detectable in wild-type C57BL/6 mice and homozygous B-hADA mice as the antibody is cross-reactive with ADA in human and mice.
