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M004043200v1
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Papers In Press, published online ahead of print June 30, 2000
J. Biol. Chem, 10.1074/jbc.M004043200
Submitted on May 11, 2000
Accepted on June 30, 2000

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. Sabatino, Clara Wong, Amanda P. Cline, Louise Pyle, Lisa J. Garrett, Patrick G. Gallagher, and David M. Bodine

Hematopoiesis Section, GMBB, NHGRI, NIH, Bethesda, MD 20892

Corresponding Author: tedyaz{at}nhgri.nih.gov

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 had 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 6/8 lines but absent or dramatically reduced levels of Ank/Agamma -globin mRNA in adult erythroid cells in 8/8 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.


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