ATP Citrate (pro-S)-Lyase / genetics* ; Animals ; Base Sequence ; Blotting, Southern ; Electrophoresis, Agar Gel ; Molecular Sequence Data ; Nucleic Acid Hybridization ; Plasmids ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Rats ; Regulatory Sequences, Nucleic Acid ; Transcription, Genetic / genetics ; Transfection
Abstract
A genomic clone, encompassing the 5' flanking region and the first seven exons of rat ATP citrate lyase gene, was isolated from a rat genomic library and sequenced. Primer-extension analysis showed that mRNA is transcribed at 4407 nucleotides upstream from the translation start site. Primer-extension analysis and sequencing of ATP citrate lyase cDNA amplified by PCR showed that the promoter used for transcription is identical in mammary gland, lung, liver, brain and kidney. Southern-blot analysis showed that the ATP citrate lyase gene exists as a single copy. The 5' flanking region contains several consensus sequences defined as promoter elements. These include a CAAT box and Sp1-binding sites. However, a TATA box lacks this promoter. The expression of the chloramphenicol acetyltransferase gene was induced by the 5' flanking region (-2370 to -1) in the CHO cell line. The 5' flanking region also contains several sequence elements that may be involved in the transcriptional regulation of the gene.