Targeting strategy
Gene targeting strategy for B-hHMMR mice. The exons 2~18 of mouse Hmmr gene that encodes the extracellular domain were replaced by human HMMR exons 2~18 in B-hHMMR mice.
Protein expression analysis
Western blot analysis of HMMR protein expression in homozygous B-hHMMR mice. Various tissue lysates were collected from wild-type C57BL/6 (+/+) mice and homozygous B-hHMMR mice (H/H), and then analyzed by western blot with species-specific anti-HMMR antibody. 30 μg total proteins were loaded for western blotting analysis. HMMR was detected in thymus.
mRNA expression analysis
Strain specific analysis of HMMR mRNA expression in wild-type C57BL/6 mice and B-hHMMR mice by RT-PCR. Thymus RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hHMMR mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human HMMR primers. Mouse Hmmr mRNA was detectable only in wild-type C57BL/6 mice. Human HMMR mRNA was detectable only in homozygous B-hHMMR mice but not in wild-type mice.
