Open AccessMolecular cloning of the murine adenosine deaminase gene from a genetically enriched source: identification and characterization of the promoter region.
Author(s) -
Diane E. Ingolia,
Muayyad R. AlUbaidi,
C Y Yeung,
H A Bigo,
David Wright,
Rodney E. Kellems
Publication year - 1986
Publication title -
molecular and cellular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.14
H-Index - 327
eISSN - 1067-8824
pISSN - 0270-7306
DOI - 10.1128/mcb.6.12.4458
Subject(s) - biology , cosmid , promoter , gene , genetics , genomic library , primer extension , microbiology and biotechnology , transcription (linguistics) , gene expression , rna , peptide sequence , linguistics , philosophy
A genomic library was prepared with DNA from a genetically enriched mouse cell line in which amplified copies of the adenosine deaminase (ADA) gene account for over 5% of the genome. Overlapping cosmid clones encompassing the entire ADA structural gene were isolated from this genomic library and used for subsequent structural and functional analyses. Nuclease protection and primer extension analyses served to identify the location of multiple transcription initiation sites at the 5' end of the structural gene. Promoter activity was found by functional analyses to reside within a 240-base-pair fragment which contains the transcription initiation sites. Sequences upstream of the transcription initiation sites are very G + C rich (77%) and include a 22 nucleotide stretch of deoxyguanylate residues and two potential Sp1 transcription factor-binding sites. Comparison of the mouse and human ADA gene promoters revealed the presence of several regions that are highly conserved with regard to both sequence content and location and may represent genetic elements which are involved in ADA gene expression.