Two phage clones, λhgACL21 and AhgACL28, harboring the 5′ flanking region of human ATP-citrate lyase (ACL) gene were identified by screening about 1.5 × 106 recombinant plaques from the EMBL3-human placental genomic DNA library. The 5′ flanking region of ACL had the CAAT box on -92 bp from the transcription initiation site (+1), however, the TATA box was not found. The primer extension and rapid amplification of eDNA end showed that mRNA is transcribed at a thymine extending 12 bp upstream of the reported eDNA end. Several consensus, sequences including four Spl binding sites, were found in the 5′ flanking region of this gene. The promoter activity was assayed by transfecting the 3′ or 5′ deletion clones of ACL-CAT plasmid into PLC/PRF5 cells. The clone that contains the part of the first intron sequences from -659 to +440 bp showed the highest CAT activity in the transient transfection assay. High promoter activities were maintained until the transcription initiation site was removed. It is suggested that the sequences from -213 to +12 which contain three Spl-binding sequences, CAAT box, and the transcription initiation site were necessary as a mean of for exerting the basal promoter activity of ACL gene.
|Original language||English (US)|
|State||Published - Dec 1 1997|
ASJC Scopus subject areas
- Molecular Biology