C57BL/6N-Rxfp1tm1(RXFP1)Bcgen/Bcgen • 113095
Gene targeting strategy for B-hRXFP1 mice. In B-hRXFP1 mice, an internal ribosome entry site (IRES2) fused to full length human RXFP1 cDNA, followed by human 3`UTR, inserted into exon 17 of the endogenous Rxfp1 gene. The human protein expression will be driven by mouse endogenous promoter, while mouse Rxfp1 gene transcription and translation will be disrupted.
Western blot analysis of RXFP1 protein expression in homozygous B-hRXFP1 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hRXFP1 mice (H/H), and then analyzed by western blot with anti-RXFP1 antibody (GeneTex, GTX108186, the antibody was cross-reactive between human and mouse). 40 μg total proteins were loaded for western blotting analysis. m/hRXFP1 was obviously detected in heart, liver and brain. m/hRXFP1 was also detected in lung and testis.
Strain specific analysis of RXFP1 mRNA expression in wild-type C57BL/6 mice and B-hRXFP1 mice by RT-PCR. Brain, lung and Heart RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hRXFP1 mice(H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human RXFP1 primers. Mouse Rxfp1 mRNA were detectable in wild-type C57BL/6 mice. Human RXFP1 mRNA was detectable only in homozygous B-hRXFP1 mice but not in wild-type mice.