Molecular cloning of the gene for the human prostaglandin transporter hPGT

Gene organization, promoter activity, and chromosomal localization

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26 Citations (Scopus)

Abstract

Prostaglandins (PGs) play diverse and important roles in human health and disease. We recently identified the first known PG transporter cDNA in the rat (PGT) and human (hPGT). To aid in the analysis of any possible human disease caused by mutations in PGT, we have cloned and characterized the hPGT gene. The gene exists as a single copy in the human genome and is comprised of 14 exons distributed over ~95 kb. Two introns disrupt putative trans-membrane spans of the coding region; each of these sites is near a highly conserved charged residue. The ~250 bp immediately 5@? to the start of exon 1 contain a TATAAA sequence (TATA box), a transcription initiation (Inr) consensus (CTCANTCT), two Sp 1 sequences (GGGCGG), and a cAMP response element (CGGCGTCA). Ligation of ~3.5 kb of 5@? flanking sequence to a luciferase reporter yielded > 15-fold activity above background when expressed in A549 human lung epithelial cells. PCR-based monochromosomal somatic cell hybrid mapping and fluorescence in situ hybridization localized hPGT to chromosome 3q21. Three microsatellites were identified, one of which was demonstrated to be polymorphic in unrelated individuals and may be useful in evaluating PGT as a candidate gene in human disease.

Original languageEnglish (US)
Pages (from-to)805-812
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume246
Issue number3
DOIs
StatePublished - May 29 1998

Fingerprint

Cloning
Molecular Cloning
Prostaglandins
Genes
Exons
TATA Box
Response Elements
Transcription
Chromosomes
Luciferases
Microsatellite Repeats
Introns
Hybrid Cells
Rats
Human Genome
Fluorescence In Situ Hybridization
Complementary DNA
Fluorescence
Health
Ligation

Keywords

  • Biological
  • Carrier proteins
  • Consensus sequence
  • CpG islands
  • Exons
  • Fluorescence
  • In situ hybridization
  • Introns
  • Microsatellite repeats
  • Molecular cloning
  • Molecular sequence data
  • Nucleic acid
  • Promoter regions
  • Prostaglandins
  • Regulatory sequences
  • Transport

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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title = "Molecular cloning of the gene for the human prostaglandin transporter hPGT: Gene organization, promoter activity, and chromosomal localization",
abstract = "Prostaglandins (PGs) play diverse and important roles in human health and disease. We recently identified the first known PG transporter cDNA in the rat (PGT) and human (hPGT). To aid in the analysis of any possible human disease caused by mutations in PGT, we have cloned and characterized the hPGT gene. The gene exists as a single copy in the human genome and is comprised of 14 exons distributed over ~95 kb. Two introns disrupt putative trans-membrane spans of the coding region; each of these sites is near a highly conserved charged residue. The ~250 bp immediately 5@? to the start of exon 1 contain a TATAAA sequence (TATA box), a transcription initiation (Inr) consensus (CTCANTCT), two Sp 1 sequences (GGGCGG), and a cAMP response element (CGGCGTCA). Ligation of ~3.5 kb of 5@? flanking sequence to a luciferase reporter yielded > 15-fold activity above background when expressed in A549 human lung epithelial cells. PCR-based monochromosomal somatic cell hybrid mapping and fluorescence in situ hybridization localized hPGT to chromosome 3q21. Three microsatellites were identified, one of which was demonstrated to be polymorphic in unrelated individuals and may be useful in evaluating PGT as a candidate gene in human disease.",
keywords = "Biological, Carrier proteins, Consensus sequence, CpG islands, Exons, Fluorescence, In situ hybridization, Introns, Microsatellite repeats, Molecular cloning, Molecular sequence data, Nucleic acid, Promoter regions, Prostaglandins, Regulatory sequences, Transport",
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T1 - Molecular cloning of the gene for the human prostaglandin transporter hPGT

T2 - Gene organization, promoter activity, and chromosomal localization

AU - Lu, Run

AU - Schuster, Victor L.

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N2 - Prostaglandins (PGs) play diverse and important roles in human health and disease. We recently identified the first known PG transporter cDNA in the rat (PGT) and human (hPGT). To aid in the analysis of any possible human disease caused by mutations in PGT, we have cloned and characterized the hPGT gene. The gene exists as a single copy in the human genome and is comprised of 14 exons distributed over ~95 kb. Two introns disrupt putative trans-membrane spans of the coding region; each of these sites is near a highly conserved charged residue. The ~250 bp immediately 5@? to the start of exon 1 contain a TATAAA sequence (TATA box), a transcription initiation (Inr) consensus (CTCANTCT), two Sp 1 sequences (GGGCGG), and a cAMP response element (CGGCGTCA). Ligation of ~3.5 kb of 5@? flanking sequence to a luciferase reporter yielded > 15-fold activity above background when expressed in A549 human lung epithelial cells. PCR-based monochromosomal somatic cell hybrid mapping and fluorescence in situ hybridization localized hPGT to chromosome 3q21. Three microsatellites were identified, one of which was demonstrated to be polymorphic in unrelated individuals and may be useful in evaluating PGT as a candidate gene in human disease.

AB - Prostaglandins (PGs) play diverse and important roles in human health and disease. We recently identified the first known PG transporter cDNA in the rat (PGT) and human (hPGT). To aid in the analysis of any possible human disease caused by mutations in PGT, we have cloned and characterized the hPGT gene. The gene exists as a single copy in the human genome and is comprised of 14 exons distributed over ~95 kb. Two introns disrupt putative trans-membrane spans of the coding region; each of these sites is near a highly conserved charged residue. The ~250 bp immediately 5@? to the start of exon 1 contain a TATAAA sequence (TATA box), a transcription initiation (Inr) consensus (CTCANTCT), two Sp 1 sequences (GGGCGG), and a cAMP response element (CGGCGTCA). Ligation of ~3.5 kb of 5@? flanking sequence to a luciferase reporter yielded > 15-fold activity above background when expressed in A549 human lung epithelial cells. PCR-based monochromosomal somatic cell hybrid mapping and fluorescence in situ hybridization localized hPGT to chromosome 3q21. Three microsatellites were identified, one of which was demonstrated to be polymorphic in unrelated individuals and may be useful in evaluating PGT as a candidate gene in human disease.

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KW - Microsatellite repeats

KW - Molecular cloning

KW - Molecular sequence data

KW - Nucleic acid

KW - Promoter regions

KW - Prostaglandins

KW - Regulatory sequences

KW - Transport

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