Nejvíce citovaný článek - PubMed ID 3020502
A reporter gene system, based on luciferase genes from Vibrio harvei, was constructed for measurement of translation nonsense suppression in Streptomyces. Using the site-directed mutagenesis the TCA codon in position 13 of the luxB gene was replaced by all of the three stop codons individually. By cloning of luxA and luxB genes under the control of strong constitutive Streptomyces promoter ermE* in plasmid pUWL201 we created Wluxl with the wild-type sequence and pWlux2, pWlux3 and pWlux4 plasmids containing TGA-, TAG- and TAA-stop codons, respectively. Streptomyces lividans TK 24 was transformed with the plasmids and the reporter system was tested by growth of the strain in the presence of streptomycin as a translation accuracy modulator. Streptomycin increased nonsense suppression on UAA nearly 10-fold and more than 20-fold on UAG. On the other hand, UGA, the most frequent stop signal in Streptomyces, the effect was negligible.
- MeSH
- bakteriální geny MeSH
- bakteriální proteiny analýza genetika MeSH
- inhibitory syntézy proteinů farmakologie MeSH
- luciferasy bakteriální analýza genetika MeSH
- methyltransferasy genetika MeSH
- mutageneze cílená MeSH
- nesmyslný kodon MeSH
- plazmidy genetika MeSH
- promotorové oblasti (genetika) MeSH
- proteosyntéza * MeSH
- reportérové geny * MeSH
- Streptomyces lividans účinky léků genetika fyziologie MeSH
- streptomycin farmakologie MeSH
- supresorové geny * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- bakteriální proteiny MeSH
- inhibitory syntézy proteinů MeSH
- luciferasy bakteriální MeSH
- methyltransferasy MeSH
- nesmyslný kodon MeSH
- rRNA (adenosine-O-2'-)methyltransferase MeSH Prohlížeč
- streptomycin MeSH
Accuracy of activity of ribosome isolated from UV-light-induced streptomycin-resistant R-21 mutant of Streptomyces granaticolor was measured in an E. coli-derived system translating poly(U) with a high rate and accuracy. Ribosomes from the R-21 mutant strain were shown to be resistant to streptomycin and about two-fold more accurate than those from the wild type. The mutant strain was found to be resistant to 1000 mg/L streptomycin (Stm) during vegetative growth while it sporulated on agar plates containing only up to 200 mg/L of Stm. The growth rate of the R-21 mutant in complex liquid medium was indistinguishable from that of the wild-type strain.
