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We have investigated the transcriptional regulation of the human embryonic zeta-globin gene promoter. First, we examined the effect that deletion of sequences 5' to zeta-globin's CCAAT box have on zeta-promoter activity in erythroid cell lines. Deletions of sequences between -116 and -556 (cap = 0) had little effect while further deletion to -84 reduced zeta-promoter activity by only 2-3-fold in both transiently and stably transfected erythroid cells. Constructs containing 67, 84 and 556 bp of zeta-globin 5' flanking region linked to a beta-galactosidase reporter gene (lacZ) and hypersensitive site -40 (HS-40) of the human alpha-globin gene cluster were then employed for the generation of transgenic mice. LacZ expression from all constructs, including a 67 bp zeta-globin promoter, was erythroid-specific and most active between 8.5 and 10.5 days post-fertilisation. By 16.5 days gestation, lacZ expression dropped 40-100-fold. These results suggest that embryonic-specific activation of the human zeta-globin promoter is conferred by a 67 bp zeta-promoter fragment containing only a CCAAT and TATA box.

Type

Journal article

Journal

Nucleic Acids Res

Publication Date

01/11/1996

Volume

24

Pages

4158 - 4164

Keywords

Animals, Cell Line, Embryo, Mammalian, Erythrocytes, Female, Fetus, Gene Expression Regulation, Developmental, Genes, Reporter, Globins, Humans, Lac Operon, Liver, Male, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Mice, Transgenic, Promoter Regions, Genetic, Transfection