Abstract
We have produced human CEA transgenic mice which were found to express CEA mRNA in all tissues. By immunoblot analysis using anti-CEA polyclonal antibody, we also detected CEA protein in all tissues. However, the molecular size of CEA in the brain was different from that in other tissues, although the mRNA size was same and no deletion nor rearrangement was detected at the DNA level. Immunohistochemical analysis of the lung and the colon showed that the expression sites were the bronchial epithelial cells of the lung and the columnar epithelial cells of the colon. Interestingly, the expression of CEA protein in the transgenic mice was polarised to the luminal side of epithelial cells similar to the normal CEA expression in human tissues. We also detected cell surface expression of human CEA on thymocytes and spleen cells and CEA expression was greatly reduced by the phosphatidylinositol-specific phospholipase C (PI-PLC) treatment.
Highlights
This fragment contains SV40 early promoter, P-globin splice site and a 2.9 kb EcoRI fragment of full length CEA cDNA but is lacking almost all of the dhfr gene
DNA solution (5 pgml-') was microinjected into the pronucleus of fertilised eggs derived from the mating of C57BL/6 or BCF1 (C57BL/6 x Balb/c) females with C57BL/6 males
The tail DNAs from these mice were screened for the presence of human CEA DNA by Southern blot analysis
Summary
This fragment contains SV40 early promoter, P-globin splice site and a 2.9 kb EcoRI fragment of full length CEA cDNA but is lacking almost all of the dhfr gene. This fragment was excised from low melting point agarose gel, purified by phenol and chloroform extraction, further purified using ELUTIP-d columns (Schleicher & Schuell GmbH: Dassel, Germany), followed by ethanol precipitation and suspended in TE (10 mM Tris pH 7.5 and 0.2 mM EDTA). DNA solution (5 pgml-') was microinjected into the pronucleus of fertilised eggs derived from the mating of C57BL/6 or BCF1 (C57BL/6 x Balb/c) females with C57BL/6 males. Eggs surviving after microinjection were reimplanted into the oviducts of pseudopregnant ddY females
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