A comparative hybridization analysis of yeast DNA with Paramecium parafusin- and different phosphoglucomutase-specific probes

E. Wyroba, Birgit H. Satir

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Molecular probes designed for the parafusin (PFUS), the Paramecium exocytic-sensitive phosphoglyco-protein, gave distinct hybridization patterns in Saccharomyces cerevisiae genomic DNA when compared with different phosphoglucomutase specific probes. These include two probes identical to segments of yeast phosphoglucomutase (PGM) genes 1 and 2. Neither of the PGM probes revealed the 7.4 and 5.9 kb fragments in Bgl II-cut yeast DNA digest detected with the 1.6 kb cloned PFUS cDNA and oligonucleotide constructed to the PFUS region (insertion 3 -1-3) not found in other species. PCR amplification with PFUS-specific primers generated yeast DNA-species of the predicted molecular size which hybridized to the I-3 probe. A search of the yeast genome database produced an unassigned nucleotide sequence that showed 55% identity to parafusin gene and 37% identity to PGM2 (the major isoform of yeast phosphoglucomutase) within the amplified region.

Original languageEnglish (US)
Pages (from-to)683-690
Number of pages8
JournalBiochemistry and Cell Biology
Volume78
Issue number6
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Phosphoglucomutase
Paramecium
Yeast
Yeasts
DNA
Genes
Molecular Probes
Oligonucleotides
Saccharomyces cerevisiae
Protein Isoforms
Complementary DNA
Amplification
Genome
Databases
Nucleotides
Polymerase Chain Reaction
Proteins

Keywords

  • Hybridization
  • Parafusin
  • PCR
  • Phosphoglucomutase
  • Yeast

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

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title = "A comparative hybridization analysis of yeast DNA with Paramecium parafusin- and different phosphoglucomutase-specific probes",
abstract = "Molecular probes designed for the parafusin (PFUS), the Paramecium exocytic-sensitive phosphoglyco-protein, gave distinct hybridization patterns in Saccharomyces cerevisiae genomic DNA when compared with different phosphoglucomutase specific probes. These include two probes identical to segments of yeast phosphoglucomutase (PGM) genes 1 and 2. Neither of the PGM probes revealed the 7.4 and 5.9 kb fragments in Bgl II-cut yeast DNA digest detected with the 1.6 kb cloned PFUS cDNA and oligonucleotide constructed to the PFUS region (insertion 3 -1-3) not found in other species. PCR amplification with PFUS-specific primers generated yeast DNA-species of the predicted molecular size which hybridized to the I-3 probe. A search of the yeast genome database produced an unassigned nucleotide sequence that showed 55{\%} identity to parafusin gene and 37{\%} identity to PGM2 (the major isoform of yeast phosphoglucomutase) within the amplified region.",
keywords = "Hybridization, Parafusin, PCR, Phosphoglucomutase, Yeast",
author = "E. Wyroba and Satir, {Birgit H.}",
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journal = "Biochemistry and Cell Biology",
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T1 - A comparative hybridization analysis of yeast DNA with Paramecium parafusin- and different phosphoglucomutase-specific probes

AU - Wyroba, E.

AU - Satir, Birgit H.

PY - 2000

Y1 - 2000

N2 - Molecular probes designed for the parafusin (PFUS), the Paramecium exocytic-sensitive phosphoglyco-protein, gave distinct hybridization patterns in Saccharomyces cerevisiae genomic DNA when compared with different phosphoglucomutase specific probes. These include two probes identical to segments of yeast phosphoglucomutase (PGM) genes 1 and 2. Neither of the PGM probes revealed the 7.4 and 5.9 kb fragments in Bgl II-cut yeast DNA digest detected with the 1.6 kb cloned PFUS cDNA and oligonucleotide constructed to the PFUS region (insertion 3 -1-3) not found in other species. PCR amplification with PFUS-specific primers generated yeast DNA-species of the predicted molecular size which hybridized to the I-3 probe. A search of the yeast genome database produced an unassigned nucleotide sequence that showed 55% identity to parafusin gene and 37% identity to PGM2 (the major isoform of yeast phosphoglucomutase) within the amplified region.

AB - Molecular probes designed for the parafusin (PFUS), the Paramecium exocytic-sensitive phosphoglyco-protein, gave distinct hybridization patterns in Saccharomyces cerevisiae genomic DNA when compared with different phosphoglucomutase specific probes. These include two probes identical to segments of yeast phosphoglucomutase (PGM) genes 1 and 2. Neither of the PGM probes revealed the 7.4 and 5.9 kb fragments in Bgl II-cut yeast DNA digest detected with the 1.6 kb cloned PFUS cDNA and oligonucleotide constructed to the PFUS region (insertion 3 -1-3) not found in other species. PCR amplification with PFUS-specific primers generated yeast DNA-species of the predicted molecular size which hybridized to the I-3 probe. A search of the yeast genome database produced an unassigned nucleotide sequence that showed 55% identity to parafusin gene and 37% identity to PGM2 (the major isoform of yeast phosphoglucomutase) within the amplified region.

KW - Hybridization

KW - Parafusin

KW - PCR

KW - Phosphoglucomutase

KW - Yeast

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