CAXII (carbonic anhydrase 12) is a type I membrane enzyme highly expressed in colon, kidney.
The exons 1~10 of mouse Car12 gene that encode signal peptide and extracellular domain are replaced by human counterparts in B-hCAXII mice. The genomic region of mouse Car12 gene that encodes transmembrane domain and cytoplasmic portion is retained. The chimeric CAXII expression is driven by endogenous mouse Car12 promoter.
CAXII protein was detected in kidney, colon, stomach (weak expression), and skin (weak expression) of homozygous B-hCAXII mice.
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mRNA expression analysis
Species specific analysis of CAXII gene expression in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCAXII mice by RT-PCR. Kidney was collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCAXII mice (H/H). Mouse Car12 mRNA was detectable only in wild-type C57BL/6JNifdc mice. Human CAXII mRNA was detectable only in homozygous B-hCAXII mice, but not in wild-type C57BL/6JNifdc mice.
Protein expression analysis
Western blot analysis of CAXII protein expression in homozygous B-hCAXII mice. Various tissue lysates were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCAXII mice (H/H), and then analyzed by western blot with species-specific anti-CAXII antibody (CST, 5864). 40 μg total proteins were loaded for western blotting analysis. The high expression of CAXII protein was detected in kidney and colon of homozygous B-hCAXII mice compared to the stomach and skin.