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Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1612-7.

Efficient studies of long-distance Bmp5 gene regulation using bacterial artificial chromosomes.

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  • 1Department of Developmental Biology, Howard Hughes Medical Institute, Beckman Center, 179 Campus Drive, Stanford, CA, 94305-5329, USA.

Abstract

The regulatory regions surrounding many genes may be large and difficult to study using standard transgenic approaches. Here we describe the use of bacterial artificial chromosome clones to rapidly survey hundreds of kilobases of DNA for potential regulatory sequences surrounding the mouse bone morphogenetic protein-5 (Bmp5) gene. Simple coinjection of large insert clones with lacZ reporter constructs recapitulates all of the sites of expression observed previously with numerous small constructs covering a large, complex regulatory region. The coinjection approach has made it possible to rapidly survey other regions of the Bmp5 gene for potential control elements, to confirm the location of several elements predicted from previous expression studies using regulatory mutations at the Bmp5 locus, to test whether Bmp5 control regions act similarly on endogenous and foreign promoters, and to show that Bmp5 control elements are capable of rescuing phenotypic effects of a Bmp5 deficiency. This rapid approach has identified new Bmp5 control regions responsible for controlling the development of specific anatomical structures in the vertebrate skeleton. A similar approach may be useful for studying complex control regions surrounding many other genes important in embryonic development and human disease.

PMID:
10677507
[PubMed - indexed for MEDLINE]
PMCID:
PMC26483
Free PMC Article
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