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Volumn 288, Issue 5474, 2000, Pages 2212-2215

Bacterial mode of replication with eukaryotic-like machinery in a hyperthermophilic archaeon

Author keywords

[No Author keywords available]

Indexed keywords

BACTERIAL DNA; URACIL DERIVATIVE;

EID: 0034705420     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5474.2212     Document Type: Article
Times cited : (195)

References (35)
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    • 14C-labeled uracil (2 μCi/ml), and labeling was continued for indicated times (Fig. 2). Agarose plugs for CHEF analyses (6.5 V/cm, a linear pulse ramp of 3 to 17 s for 20 hours, 14°C) were prepared from 5-ml cultures by using a modification of standard protocols. Gels and filters (1 to 2 weeks of exposure to Storage Phosphor screens) were analyzed with a Storm imaging system (Molecular Dynamics, Sunnyvale, CA). Radioactivity of each Not I fragment was determined by using ImageQuant software and following the manufacturer's recommendations. To ensure that the results obtained were not an artifact of the puromycin treatment, the relative steady-state levels of Not I fragments were also investigated in untreated early exponential stage cultures. In this case, CHEF gels were stained with VistraGreen (Molecular Dynamics), a DNA-specific fluorescence dye, and were directly scanned with a blue fluorescence mode of a Storm system. Relative frequencies for fragments were obtained by dividing the amount of radioactivity/fluorescence of a given band by its size calculated on the basis of the determined genome sequence. All analyzed signals were within a linear range of a Storm system. The given values were normalized to the minimal value of one arbitrary unit and are averages of two independent measurements.
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    • This work was supported by research grants from the European Union (BIO-CT 96-0488) and the Association de Recherche contre le Cancer. H.M. acknowledges financial support from the European Union and thanks U. Liebl for helpful discussions. We thank J. Weissenbach and his staff for sequencing the complete genome of P. abyssi.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.