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  • 1
    Electronic Resource
    Electronic Resource
    Positive selection for Dictyostelium mutants lacking uridine monophosphate synthase activity based on resistance to 5-fluoro-orotic acid (1990)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 396-402 
    Details
    ISSN: 0192-253X
    Keywords: Transformation ; Dictyostelium discoideum ; UMP-synthase/ura-complementation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Whereas transformation ofDictyostelium discoideum now can be done routinely and reliably, there is increasing demand for a system that allows selection of cells that have gone through repeated transformation cycles. Such a system is presented here. Selection is based on resistance to 5-fluoro-orotic-acid (5-FOA). D. discoi-deum is highly sensitive to this drug, and cells can survive only in the presence of 5-FOA if they carry a defective UMP-synthase gene. After transformation with a plasmid carrying a cloned version of the UMP-synthase gene, 5-FOA resistant cells are obtained at high frequency. Because 5-FOA-resistant cells depend on exogenously added uracil, these cells can be taken through a second round of transformation. A plasmid-borne UMP-synthase gene renders 5-FOA-resistant cells phenotypically ura +. Finally, cells can be further transformed using the standard G418 selection system.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020110512
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  • 2
    Electronic Resource
    Electronic Resource
    Nonsense suppression in Dictyostelium discoideum (1990)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 410-417 
    Details
    ISSN: 0192-253X
    Keywords: Suppressor †RNA genes ; yeast ; lacZ expression in D. discoideum ; †RNATrp genes ; †RNAGlu genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We describe the generation of Dictyostelium discoideum cell lines that carry different suppressor †RNA genes. These genes were constructed by primer-directed mutagenesis changing a †RNATrp(CCA) gene from D. discoideum to a †RNATrp(amber) gene and changing a †RNAGlu(UUC) gene from D. discoideum to a †RNAGlu(ochre) as well as a †RNAGlu(amber) gene. These genes were stably integrated into the D. discoideum genome together with a reporter gene. An actin 6::lacZ gene fusion carrying corresponding translational stop signals served as a reported. Active β-galactosidase is expressed only in D. discoideum strains that contain, in addition to the reporter, a functional suppressor †RNA. Both amber suppressors are active in D. discoideum without interfering significantly with cell growth and development. We failed, however, to establish cell lines containing a functional †RNAGlu(ochre) suppressor. This may be due to the fact that nearly every message from D. discoideum known so far terminates with UAA. Therefore a †RNA capable of reading this termination codon may not be compatible with cell growth.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020110514
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    Paper (German National Licenses)
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