Basic Information
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Targeting strategy
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Gene targeting strategy for B-hAOC3 mice.
Exons 1~4 of the mouse Aoc3 gene, which encodes the extracellular domain, were replaced by human AOC3 exons 1~4 in B-hAOC3 mice.
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mRNA expression analysis
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Strain specific analysis of AOC3 mRNA expression in wild-type C57BL/6N mice and B-hAOC3 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hAOC3 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human AOC3 primers. Mouse Aoc3 mRNA was detectable only in wild-type C57BL/6N mice. Human AOC3 mRNA was detectable only in homozygous B-hAOC3 mice.
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Protein expression analysis
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Western blot analysis of AOC3 protein expression in homozygous B-hAOC3 mice. Various tissue lysates were collected from wild-type C57BL/6N (+/+) mice and homozygous B-hAOC3 mice (H/H), and then analyzed by western blot with anti-AOC3 antibody. 40μg total proteins were loaded for western blotting analysis. AOC3 was detected in heart, spleen, kidney, lung and stomach of wild-type C57BL/6N mice and spleen, lung and stomach in homozygous B-hAOC3 mice.
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Summary
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mRNA expression analysis:
Mouse Aoc3 mRNA was detectable in lung of wild-type C57BL/6N mice. Human AOC3 mRNA was detectable in B-hAOC3 mice.
Protein expression analysis:
Human and mouse AOC3 were detectable in spleen, lung and stomach both in homozygous B-hAOC3 mice and wild-type C57BL/6N mice, as the antibody cross-recognizes both human and mouse AOC3.
