Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCTSG mice. The exon 1-5 of mouse Ctsg gene that encodes the full-length protein was replaced by human CTSG exon 1-5 in B-hCTSG mice.
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Protein expression analysis
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Western blot analysis of CTSG protein expression in homozygous B-hCTSG mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCTSG mice (H/H), and then analyzed by western blot with species-specific anti-CTSG antibody. 20 μg total proteins were loaded for western blotting analysis. hCTSG was detected in BM from homozygous mice but not in spleen, thymus,liver and kidney.
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mRNA expression analysis
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Strain specific analysis of CTSG mRNA expression in wild-type C57BL/6 mice and B-hCTSG mice by RT-PCR. BM RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hCTSG mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CTSG primers. Mouse CTSG mRNA was detectable only in wild-type C57BL/6 mice. Human CTSG mRNA was detectable only in homozygous B-hCTSG mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Mouse Ctsg mRNA was detectable in BM of wild-type C57BL/6 mice. Human CTSG mRNA was detectable in B-hCTSG mice.
Protein expression analysis:
Human CTSG was detectable in BM of homozygous B-hCTSG mice.