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Originally published In Press as doi:10.1074/jbc.M004043200 on June 30, 2000

J. Biol. Chem., Vol. 275, Issue 37, 28549-28554, September 15, 2000
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A Minimal Ankyrin Promoter Linked to a Human gamma -Globin Gene Demonstrates Erythroid Specific Copy Number Dependent Expression with Minimal Position or Enhancer Dependence in Transgenic Mice*

Denise E. SabatinoDagger §, Clara Wong, Amanda P. ClineDagger , Louise PyleDagger , Lisa J. Garrett||, Patrick G. Gallagher, and David M. BodineDagger **

From the Dagger  Hematopoiesis Section, Genetics and Molecular Biology Branch, NHGRI, National Institutes of Health, Bethesda, Maryland 20892, the § Graduate Genetics Program, The George Washington University, Washington, DC 20052, the  Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06520, and the || Genetic Disease Research Branch, NHGRI, National Institutes of Health, Bethesda, Maryland 20892

In red blood cells ankyrin (ANK-1) provides the primary linkage between the erythrocyte membrane skeleton and the plasma membrane. We have previously demonstrated that a 271-bp 5'-flanking region of the ANK-1 gene has promoter activity in erythroid, but not non-erythroid, cell lines. To determine whether the ankyrin promoter could direct erythroid-specific expression in vivo, we analyzed transgenic mice containing the ankyrin promoter fused to the human Agamma -globin gene. Sixteen of 17 lines expressed the transgene in erythroid cells indicating nearly position-independent expression. We also observed a significant correlation between the level of Ank/Agamma -globin mRNA and transgene copy number. The level of Ank/Agamma mRNA averaged 11% of mouse alpha -globin mRNA per gene copy at all developmental stages. The addition of the HS2 enhancer from the beta -globin locus control region to the Ank/Agamma -globin transgene resulted in Ank/Agamma -globin mRNA expression in embryonic and fetal erythroid cells in six of eight lines but resulted in absent or dramatically reduced levels of Ank/Agamma -globin mRNA in adult erythroid cells in eight of eight transgenic lines. These data indicate that the minimal ankyrin promoter contains all sequences necessary and sufficient for erythroid-specific, copy number-dependent, position-independent expression of the human Agamma -globin gene.

* This work was supported by grants from the National Institutes of Health and The March of Dimes Birth Defects Foundation (to P. G. G.).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.

** To whom correspondence should be addressed: Hematopoiesis Section, Genetics and Molecular Biology Branch, NHGRI, National Institutes of Health, Building 49, Room 3A14 MSC 4442, Bethesda, MD 20892-4442. Tel.: 301-402-0902; Fax: 301-402-4929; E-mail: tedyaz@nhgri.nih.gov.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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