Gene targeting strategy for B-hTEM7R mice. A chimeric CDS that will encode the signal peptide, extracellular domain of human TEM7R and transmembrane, cytoplasmic domain of mouse TEM7R replaced the coding sequence of mouse exon1. The targeted mice will express the chimeric TEM7R protein, while mouse TEM7R will no longer express.

Strain specific analysis of TEM7R mRNA expression in wild-type C57BL/6 mice and B-hTEM7R mice by RT-PCR. Lung and kidney RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hTEM7R mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TEM7R primers. Mouse Tem7r mRNA was only detectable in wild-type C57BL/6 mice. Human TEM7R mRNA was only detectable in homozygous B-hTEM7R mice but not in wild-type mice.

Western blot analysis of TEM7R protein expression in homozygous B-hTEM7R mice. Various tissue lysates were collected from wild-type C57BL/6N (+/+) mice and homozygous B-hTEM7R mice (H/H), and then analyzed by western blot with anti-TEM7R antibody. 30 μg total proteins were loaded for western blotting analysis. TEM7R was detected in kidney, bladder and cerebellum.