Basic Information
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Targeting strategy
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Gene targeting strategy for B-hMELTF mice.
The exons 2~16 of mouse Meltf gene that encode the full-length protein except the signal peptide were replaced by human MELTF exons 2~16 in B-hMELTF mice. -
Protein expression analysis
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Western blot analysis of MELTF protein expression in homozygous B-hMELTF mice. Various tissue lysates were collected from C57BL/6 wildtype mice (+/+) and homozygous B-hMELTF mice (H/H), and then analyzed by western blot with anti-MELTF antibody. 40 μg total proteins were loaded for western blotting analysis. MELTF was detected in testis of wildtype mice (+/+) and homozygous B-hMELTF mice (H/H), as the antibody is crossly reactive with human and mice.
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mRNA expression analysis
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Strain specific analysis of MELTF mRNA expression in wild-type C57BL/6 mice and B-hMELTF mice by RT-PCR. Brain and kidney RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hMELTF mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human MELTF primers. Mouse Meltf mRNA was detectable only in wild-type C57BL/6 mice. Human MELTF mRNA was detectable only in homozygous B-hMELTF mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Mouse Meltf mRNA was detectable in brain and kidney of wild-type C57BL/6 mice. Human MELTF mRNA was detectable in B-hMELTF mice.Protein expression analysis:
Human MELTF was detected in testis of wildtype mice and homozygous B-hMELTF mice, as the antibody is crossly reactive with human and mice.
