Cloning and functional analysis of the swine eNOS promoter

Kimberly M. Shontz, Bi Zhou, C. Yung Yu, Baogen Y. Su

Research output: Contribution to journalArticlepeer-review

Abstract

We have cloned the swine eNOS promoter and analyzed its function in newborn swine pulmonary artery endothelial cells (PAECs). Analysis of the 2.1 kb 5' flanking region revealed that the swine eNOS promoter is, like its counterparts in human and other species, a TATA-less promoter. The transcription start site, determined by 5' RLM-RACE, was located 62 bp upstream of the translation start codon. Promoter activity was demonstrated by transient transfection of 5' deletion promoter/luciferase constructs into swine PAECs, and indicated that the proximal region from - 227 to - 82 was necessary for basal promoter activity. Positive cis-regulatory elements were present from - 227 to - 1290, while negative cis-regulatory elements may be present from - 1290 to - 1926 bp. Electrophoretic mobility shift assay (EMSA) of the proximal region demonstrated that multiprotein complexes were formed in the conserved proximal region of the swine eNOS promoter and a novel Spl site at - 68/ - 59 was involved in the formation of these complexes.

Original languageEnglish (US)
Pages (from-to)62-67
Number of pages6
JournalDNA Sequence - Journal of DNA Sequencing and Mapping
Volume19
Issue number1
DOIs
StatePublished - 2008

Keywords

  • 5' RLM-RACE
  • Cis-regulatory elements
  • Pulmonary endothelial cells
  • Swine eNOS promoter

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics
  • Endocrinology

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