Gene targeting strategy for B-hITGA7 mice. The exons 2-24 of mouse Itga7 gene that extracellular domain are replaced by human counterparts in B-hITGA7 mice. The genomic region of mouse ITGA7 gene that encodes cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric ITGA7 expression is driven by endogenous mouse Itga7 promoter, while mouse Itga7 gene transcription and translation will be disrupted.

Western blot analysis of ITGA7 protein expression in homozygous B-hITGA7 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hITGA7 mice (H/H), and then analyzed by western blot with anti-ITGA7 antibody(scbt, sc-515716, cross-reactive between human and mouse). 40 μg total proteins were loaded for western blotting analysis.

Note:Under reducing conditions, the radiolabeled fragment of integrin α7 behaved as a protein of 97 kDa. doi.org/10.1074/jbc.270.16.9227

Strain specific analysis of ITGA7 mRNA expression in wild-type C57BL/6 mice and B-hITGA7 mice by RT-PCR. Stomach RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hITGA7 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human ITGA7 primers. Mouse Itga7 mRNA was detectable only in wild-type C57BL/6 mice. Human ITGA7 mRNA was detectable only in homozygous B-hITGA7 mice but not in wild-type mice.