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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