Nielsen, Lars B. and McCormick, Sally P. A. and Pierotti, Vincenzo and Tam, Carmen and Gunn, Michael D. and Shizuya, Hiroaki and Young, Stephen G. (1997) Human Apolipoprotein B Transgenic Mice Generated with 207- and 145-Kilobase Pair Bacterial Artificial Chromosomes. Evidence that a distant 5'-element confers appropriate transgene expression in the intestine. Journal of Biological Chemistry, 272 (47). pp. 29752-29758. ISSN 0021-9258 http://resolver.caltech.edu/CaltechAUTHORS:NIEjbc97
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We reported previously that ~80-kilobase pair (kb) P1 bacteriophage clones spanning either the human or mouse apoB gene (clones p158 and p649, respectively) confer apoB expression in the liver of transgenic mice, but not in the intestine. We hypothesized that the absence of intestinal expression was due to the fact that these clones lacked a distant DNA element controlling intestinal expression. To test this possibility, transgenic mice were generated with 145- and 207-kb bacterial artificial chromosomes (BACs) that contained the human apoB gene and more extensive 5'- and 3'-flanking sequences. RNase protection, in situ hybridization, immunohistochemical, and genetic complementation studies revealed that the BAC transgenic mice manifested appropriate apoB gene expression in both the intestine and the liver, indicating that both BACs contained the distant intestinal element. To determine whether the regulatory element was located 5' or 3' to the apoB gene, transgenic mice were generated by co-microinjecting embryos with p158 and either the 5'- or 3'-sequences from the 145-kb BAC. Analysis of these mice indicated that the apoB gene's intestinal element is located 5' to the structural gene. Cumulatively, the transgenic mouse studies suggest that the intestinal element is located between -33 and -70 kb 5' to the apoB gene.
|Additional Information:||©1997 by The American Society for Biochemistry and Molecular Biology, Inc. (Received for publication, May 23, 1997, and in revised form, August 22, 1997) We thank J. Ng for help with mouse breeding, L. Prentice for preparing tissue sections for in situ hybridization, L. Flynn for the rabbit antiserum to mouse apoB, J. Carroll and A. Corder for graphics, and S. Ordway and G. Howard for editorial assistance. This work was supported in part by National Institutes of Health Grant HL47660; fellowship awards from the American Heart Association, California Affiliate (to L.B.N. and S.P.A.M.); and a travel grant from the Danish Heart Association/Medical Research Council (to L.B.N.). The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.|
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