Isolation and characterization of a cdc 2 cDNA from Dictyostelium discoideum.
A cdc2 homologous sequence was amplified from Dictyostelium discoideum by the polymerase chain reaction and used to isolate several cDNA clones. The amino acid sequence encoded by these cDNAs exhibited approx. 60% identity to the Cdc2 proteins of other species. A cDNA containing the entire coding sequence complemented the temperature sensitive cdc28 mutant of Saccharomyces cerevisiae, although growth of the transformants was slow and limited. Southern blot analysis of restriction digests under high stringency conditions provided evidence that Dictyostelium contains a single cdc2 gene, although at lower stringency multiple fragments were detected, suggesting the existence of a cdc2 gene family. Northern blot analysis of RNA from different stages of Dictyostelium development showed that cdc2 mRNA levels increased during aggregation and then decreased to low levels by the pseudoplasmodial stage of development. By contrast, cdc2 mRNA levels remained relatively constant as cells passed from exponential growth to the stationary phase.
Michaelis
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Department of Microbiology, University of British Columbia, Vancouver, Canada.
Weeks
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D016203:CDC2 Protein Kinase / Q000235:genetics*; D002454:Cell Differentiation; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D004023:Dictyostelium / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D012331:RNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D012689:Sequence Homology, Nucleic Acid; D013696:Temperature
D004271:DNA, Fungal; D009838:Oligodeoxyribonucleotides; D012331:RNA, Fungal; D012333:RNA, Messenger; D016203:CDC2 Protein Kinase