Cloning and characterization of the 5′ flanking region of human ATP-citrate lyase gene

S. W. Park, Y. A. Moon, K. S. Kim, Y. H. Ahn, Y. S. Kim

Research output: Contribution to journalArticle

Abstract

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 languageEnglish (US)
JournalFASEB Journal
Volume11
Issue number9
StatePublished - 1997

Fingerprint

ATP Citrate (pro-S)-Lyase
ATP citrate synthase
5' Flanking Region
Cloning
Organism Cloning
molecular cloning
Transcription Initiation Site
Genes
transcription (genetics)
Clone Cells
promoter regions
clones
genes
TATA box
thymine
TATA Box
Bacteriophages
Thymine
Genomic Library
DNA libraries

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Park, S. W., Moon, Y. A., Kim, K. S., Ahn, Y. H., & Kim, Y. S. (1997). Cloning and characterization of the 5′ flanking region of human ATP-citrate lyase gene. FASEB Journal, 11(9).

Cloning and characterization of the 5′ flanking region of human ATP-citrate lyase gene. / Park, S. W.; Moon, Y. A.; Kim, K. S.; Ahn, Y. H.; Kim, Y. S.

In: FASEB Journal, Vol. 11, No. 9, 1997.

Research output: Contribution to journalArticle

Park, SW, Moon, YA, Kim, KS, Ahn, YH & Kim, YS 1997, 'Cloning and characterization of the 5′ flanking region of human ATP-citrate lyase gene', FASEB Journal, vol. 11, no. 9.
Park, S. W. ; Moon, Y. A. ; Kim, K. S. ; Ahn, Y. H. ; Kim, Y. S. / Cloning and characterization of the 5′ flanking region of human ATP-citrate lyase gene. In: FASEB Journal. 1997 ; Vol. 11, No. 9.
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