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Efficient studies of long-distance Bmp5 gene regulation using bacterial artificial chromosomes
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| Zeitschriftentitel: | Proceedings of the National Academy of Sciences |
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| Personen und Körperschaften: | , , , |
| In: | Proceedings of the National Academy of Sciences, 97, 2000, 4, S. 1612-1617 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Proceedings of the National Academy of Sciences
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| Schlagwörter: |
| Zusammenfassung: | <jats:p> 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 <jats:italic>bone morphogenetic protein-5</jats:italic> ( <jats:italic>Bmp5</jats:italic> ) gene. Simple coinjection of large insert clones with <jats:italic>lacZ</jats:italic> 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 <jats:italic>Bmp5</jats:italic> gene for potential control elements, to confirm the location of several elements predicted from previous expression studies using regulatory mutations at the <jats:italic>Bmp5</jats:italic> locus, to test whether <jats:italic>Bmp5</jats:italic> control regions act similarly on endogenous and foreign promoters, and to show that <jats:italic>Bmp5</jats:italic> control elements are capable of rescuing phenotypic effects of a <jats:italic>Bmp5</jats:italic> deficiency. This rapid approach has identified new <jats:italic>Bmp5</jats:italic> 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. </jats:p> |
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| Umfang: | 1612-1617 |
| ISSN: |
0027-8424
1091-6490 |
| DOI: | 10.1073/pnas.97.4.1612 |