Abstract:
:Mutations were introduced in 7 kilobases of 5'-flanking rat alpha 1-fetoprotein (AFP) genomic DNA, linked to the chloramphenicol acetyltransferase gene. AFP promoter activity and its repression by a glucocorticoid hormone were assessed by stable and transient expression assays. Stable transfection assays were more sensitive and accurate than transient expression assays in a Morris 7777 rat hepatoma recipient (Hepa7.6), selected for its strong AFP repression by dexamethasone. The segment of DNA encompassing a hepatocyte-constitutive chromatin DNase I-hypersensitive site at -3.7 kilobases and a liver developmental stage-specific site at -2.5 kilobases contains interacting enhancer elements sufficient for high AFP promoter activity in Hepa7.6 or HepG2 cells. Deletions and point mutations define an upstream promoter domain of AFP gene activation, operating with at least three distinct promoter-activating elements, PEI at -65 base pairs, PEII at -120 base pairs, and DE at -160 base pairs. PEI and PEII share homologies with albumin promoter sequences, PEII is a near-consensus nuclear factor I recognition sequence, and DE overlaps a glucocorticoid receptor recognition sequence. An element conferring glucocorticoid repression of AFP gene activity is located in the upstream AFP promoter domain. Receptor-binding assays indicate that this element is the glucocorticoid receptor recognition sequence which overlaps with promoter-activating element DE.
journal_name
Mol Cell Bioljournal_title
Molecular and cellular biologyauthors
Guertin M,LaRue H,Bernier D,Wrange O,Chevrette M,Gingras MC,Bélanger Ldoi
10.1128/mcb.8.4.1398subject
Has Abstractpub_date
1988-04-01 00:00:00pages
1398-407issue
4eissn
0270-7306issn
1098-5549journal_volume
8pub_type
杂志文章abstract::Site-specific recombination provides a powerful tool for studying gene function at predetermined chromosomal sites. Here we describe the use of a blasticidin resistance system to select for recombination in mammalian cells using the yeast enzyme FLP. The vector is designed so that site-specific recombination reconstru...
journal_title:Molecular and cellular biology
pub_type: 杂志文章
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更新日期:1992-04-01 00:00:00
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pub_type: 杂志文章
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更新日期:1993-01-01 00:00:00
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pub_type: 杂志文章
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更新日期:1994-09-01 00:00:00
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更新日期:2012-12-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/mcb.18.12.7235
更新日期:1998-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:1992-05-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章,收录出版
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更新日期:2013-11-01 00:00:00
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journal_title:Molecular and cellular biology
pub_type: 杂志文章
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更新日期:1984-08-01 00:00:00
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pub_type: 杂志文章
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更新日期:2001-08-01 00:00:00