Basic Information
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Targeting strategy
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Gene targeting strategy for B-hORAI1 mice.
The exon 2 of mouse Orai1 gene that encodes the extracellular domain was replaced by human ORAI1 exon 2 in B-hORAI1 mice. -
Protein expression analysis
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Western blot analysis of ORAI1 protein expression in homozygous B-hORAI1 mice. Various tissue lysates were collected from C57BL/6 wildtype mice (+/+) and homozygous B-hORAI1 mice (H/H) , and then analyzed by western blot with species-specific anti-ORAI1 antibody. 32 μg total proteins were loaded for western blotting analysis. Human ORAI1 was detected in spleen/lung/kidney/liver and small intestine of wildtype mice (+/+) and homozygous B-hORAI1 mice (H/H), as the antibody is partly crossly reactive with human and mice.
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mRNA expression analysis
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Strain specific analysis of ORAI1 mRNA expression in wild-type C57BL/6 mice and B-hORAI1 mice by RT-PCR. Spleen and lung RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hORAI1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human ORAI1 primers. Mouse Orai1 mRNA was detectable only in wild-type C57BL/6 mice. Human ORAI1 mRNA was detectable only in homozygous B-hORAI1 mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Mouse Orai1 mRNA was detectable in spleen and lung of wild-type C57BL/6 mice. Human ORAI1 mRNA was detectable in B-hORAI1 mice.Protein expression analysis:
Human ORAI1 was detected in spleen/lung/kidney/liver/small intestine of wildtype mice and homozygous B-hORAI1 mice, as the antibody is partly crossly reactive with human and mice.
