A comparative hybridization analysis of yeast DNA with Paramecium parafusin- and different phosphoglucomutase-specific probes.
Molecular probes designed for the parafusin (PFUS), the Paramecium exocytic-sensitive phosphoglycoprotein, 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--I-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.
Wyroba
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Department of Cell Biology, Nencki Institute of Experimental Biology, Warsaw, Poland. wyroba@nencki.gov.pl
Satir
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BH
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D018076:DNA, Complementary; D016681:Genome, Fungal; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010247:Paramecium / Q000235:genetics*; D010733:Phosphoglucomutase / Q000235:genetics; D010750:Phosphoproteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D016415:Sequence Alignment
D018076:DNA, Complementary; D015345:Oligonucleotide Probes; D010750:Phosphoproteins; D015800:Protozoan Proteins; C087273:parafusin protein, Paramecium; D010733:Phosphoglucomutase