Gene targeting strategy for B-hGHRL mice. The exons 2~5 of mouse Ghrl gene that encodes the full-length protein were replaced by human GHRL exons 3~6 in B-hGHRL mice.

Strain specific Obestatin & Ghrelin expression analysis in homozygous B-hGHRL mice by ELISA. Serum was collected from wild-type mice (+/+) and homozygous B-hGHRL mice (H/H), and analyzed by ELISA with Obestatin & Ghrelin ELISA kit. Obestatin was detectable both in wild-type mice and homozygous B-hGHRL mice, as the ELISA kit is cross-recognize both human and mouse Obestatin. Human Ghrelin was detected in homozygous B-hGHRL mice but not in wild-type mice.

Strain specific analysis of GHRL mRNA expression in wild-type C57BL/6 mice and B-hGHRL mice by RT-PCR. Brain RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hGHRL mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with human GHRL primers. Human GHRL mRNA was detected in homozygous B-hGHRL mice but not in wild-type mice.