Gene targeting strategy for B-hGRP mice.
The exons 1-3 of mouse Grp gene that encode the full-length protein were replaced by human GRP exons 1-3 in B-hGRP mice.
mRNA and Protein expression analysis
Strain specific analysis of GRP gene expression in wild-type mice and B-hGRP mice by RT-PCR. Mouse Grp mRNA was detectable in brain of wild-type mice (+/+). Human GRP mRNA was detectable only in homozygous B-hGRP but not in wild-type mice.
Strain specific GRP expression analysis in homozygous B-hGRP mice by western blot. Brain tissue was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hGRP mice (H/H), and analyzed by western blot with anti-GRP antibody. GRP was detectable in wild type mice and homozygous B-hGRP mice, as the antibody is crossly reactive with GRP in human and mice.
- mRNA expression analysis:
Mouse Grp mRNA was detectable in brain of wild-type (+/+). Human GRP mRNA was detectable in B-hGRP mice (H/H) but not in wild-type mice.
- Protein expression analysis:
Human GRP was exclusively detectable in homozygous B-hGRP mice.