Gene targeting strategy for B-Tg(hCEACAM3-hCEACAM5-hCEACAM6-hCEACAM7) mice. Bacterial artificial chromosome (BAC) clone CH17-137P16 for the human CEACAM3, CEACAM5, CEACAM6 and CEACAM7 genes were integrated into the mouse genome randomly.

Strain specific analysis of CEACAM mRNA expression in wild-type C57BL/6N mice and B-Tg(hCEACAM3-hCEACAM5-hCEACAM6-hCEACAM7) mice by RT-PCR. Large intestine RNA were isolated from wild-type C57BL/6N mice (+/+) and transgenic B-Tg(hCEACAM3-hCEACAM5-hCEACAM6-hCEACAM7) mice, then cDNA libraries were synthesized by reverse transcription, followed by PCR with human CEACAM primers. Human CEACAM5 and CEACAM6 were only detectable in transgenic mice, but not in wild-type mice.

Western blot analysis of CEACAM5 protein expression in B-Tg(hCEACAM3-hCEACAM5-hCEACAM6-hCEACAM7) mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and B-Tg(hCEACAM3-hCEACAM5-hCEACAM6-hCEACAM7) mice, and then analyzed by western blot with anti-CEACAM5 antibody (Invitrogen, PA5-88533). 40μg total proteins were loaded for western blotting analysis. Human CEACAM5 was only detectable in colon, small intestine, stomach and lung in transgenic mice.

Western blot analysis of CEACAM5 protein expression in B-Tg(hCEACAM3-hCEACAM5-hCEACAM6-hCEACAM7) mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and B-Tg(hCEACAM3-hCEACAM5-hCEACAM6-hCEACAM7) mice, and then analyzed by western blot with anti-CEACAM6 antibody (abcam, ab275022). 40 μg total proteins were loaded for western blotting analysis. CEACAM6 was only detectable in lung, large intestine and spleen in transgenic mice, but not in wild-type mice.