Thymidine uptake, thymidine incorporation, and thymidine kinase activity in marine bacterium isolates

Jeffrey, Wade H.; Paul, John H.

doi:10.1128/aem.56.5.1367-1372.1990

Cited by 34 publications

(16 citation statements)

References 42 publications

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“…The thymidine incorporation rates for species of Clostridium and Bacillus (Table 3) are consistent with recent literature values, although they are near the bottom of the range. Published thymidine incorporation rates for aerobic bacterial cultures lie in the range 0.1-771.7 × 10 -21 mol cell -1 h -1 [5,8,[30][31][32] and for anaerobes 0.3-21.0× 10 -21 mol cell -1 h -1 [6]. Incorporation rates per cell for data from this study yield a range of 1.8-5.0 × 10 -21 mol cell -1 h -1, calculated on the basis of cell numbers at the beginning of thymidine incorporation [5].…”

Section: Discussionmentioning

confidence: 99%

Incorporation of [methyl-3H]thymidine by obligate and facultative anaerobic bacteria when grown under defined culture conditions

Wellsbury

1993

FEMS Microbiology Ecology

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Incorporation of [methyl-3H]thymidine into bacterial DNA was determined for a range of axenic anaerobic bacterial cultures: fermentative heterotrophs, sulphate-reducing bacteria, purple sulphur bacteria, acetogens and methanogens. Anaerobically growing Bacillus sp. and the obligate aerobe Thiobacillus ferrooxidans were also investigated. Actively growing cultures of sulphate-reducing bacteria belonging to the genera Desulfovibrio, Desulfotomaculum, Desulfobacter, Desulfobotulus and Desulfobulbus, purple sulphur bacteria (Chromatium vinosum OP2 and Thiocapsa roseopersicina OP1), methanogens (Methanococcus GS16 and Methanosarcina barkeri) and an acetogen (Acetobacterium woodii) did not incorporate [methyl-3H]thymidine into DNA. The only obligate anaerobes in which thymidine incorporation into DNA could be unequivocally demonstrated were members of the genus Clostridium. Anaerobically growing Bacillus sp. also incorporated thymidine. These data demonstrate that pure culturerepresentatives of major groups of anaerobic bacteria involved in the terminal oxidation of organic carbon and anoxygenic phototrophs within sediments are unable to incorporate [methyl-3H]thymidine into DNA, although some obligate and facultative anaerobes can. Variability in thymidine incorporation amongst pure culture isolates indicates that unless existing techniques can be calibrated to take this into consideration then productivity estimates in both aerobic and anaerobic environments may be greatly underestimated using the [methyl-3H]thymidine technique.

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Section: Discussionmentioning

confidence: 99%

Incorporation of [methyl-3H]thymidine by obligate and facultative anaerobic bacteria when grown under defined culture conditions

Wellsbury

1993

FEMS Microbiology Ecology

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“…Furthermore, when pre-incubating the enzyme at 0°C for one hour, the measured activity at 21°C was 10-fold lower, while pre-incubation at 37°C for one hour resulted in irreversible denaturation. These data can be important for the interpretation of the results obtained in the past studies, when the indirect activity of TK1 from aquatic bacteria was measured at 37°C (Jeffrey & Paul, 1990). In short, when measuring the activity of bacteria isolated from cold niches by the incorporation of 3 H-dT into newly synthesized DNA, one should keep in mind that the activity should be measured at different temperatures.…”

Section: Discussionmentioning

confidence: 98%

Deoxyribonucleoside kinases in two aquatic bacteria with high specificity for thymidine and deoxyadenosine

Tinta¹,

Christiansen

Konrad

et al. 2012

FEMS Microbiol Lett

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Deoxyribonucleoside kinases (dNKs) are essential in the mammalian cell but their ‘importance’ in bacteria, especially aquatic ones, is less clear. We studied two aquatic bacteria, Gram‐negative Flavobacterium psychrophilum JIP02/86 and Polaribacter sp. MED152, for their ability to salvage deoxyribonucleosides (dNs). Both had a Gram‐positive‐type thymidine kinase (TK1), which could phosphorylate thymidine, and one non‐TK1 dNK, which could efficiently phosphorylate deoxyadenosine and slightly also deoxycytosine. Surprisingly, the four tested dNKs could not phosphorylate deoxyguanosine, and apparently, these two bacteria are missing this activity. When tens of available aquatic bacteria genomes were examined for the presence of dNKs, a majority had at least a TK1‐like gene, but several lacked any dNKs. Apparently, among aquatic bacteria, the role of the dN salvage varies.

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“…Both these plasmids encode for resistance to the antibiotics kanamycin and streptomycin. pGQ3 is a derivative of pKT230 (3) that contains the Escherichia coli thymidine kinase gene (7,16). pKT230 is a derivative of RSF1010 and pACYC177 (3).…”

Section: Methodsmentioning

confidence: 99%

Natural plasmid transformation in a high-frequency-of-transformation marine Vibrio strain

Frischer

Thurmond

Paul

1990

Appl Environ Microbiol

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The estuarine bacterium Vibrio strain DI-9 has been shown to be naturally transformable with both broad host range plasmid multimers and homologous chromosomal DNA at average frequencies of 3.5 x 10-9 and 3.4 x 1O' transformants per recipient, respectively. Growth of plasmid transformants in nonselective medium resulted in cured strains that transformed 6 to 42,857 times more frequently than the parental strain, depending on the type of transforming DNA. These high-frequency-of-transformation (Hff) strains were transformed at frequencies ranging from 1.1 x 10-8 to 1.3 x 1O-4 transformants per recipient with plasmid DNA and at an average frequency of 8.3 x 10-5 transformants per recipient with homologous chromosomal DNA. The highest transformation frequencies were observed by using multimers of an R1162 derivative carrying the transposon Tn5 (pQSR50). Probing of total DNA preparations from one of the cured strains demonstrated that no plasmid DNA remained in the cured strains which may have provided homology to the transforming DNA. Al transformants and cured strains could be differentiated from the parental strains by colony morphology. DNA binding studies indicated that late-log-phase Hff strains bound [31H]bacteriophage lambda DNA 2.1 times more rapidly than the parental strain. These results suggest that the original plasmid transformation event of strain DI-9 was the result of uptake and expression of plasmid DNA by a competent mutant (Hff strain). Additionally, it was found that a strain of Vibrio parahaemolyticus, USFS 3420, could be naturally transformed with plasmid DNA. Natural plasmid transformation by high-transforming mutants may be a means of plasmid acquisition by natural aquatic bacterial populations.

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Thymidine uptake, thymidine incorporation, and thymidine kinase activity in marine bacterium isolates

Cited by 34 publications

References 42 publications

Incorporation of [methyl-3H]thymidine by obligate and facultative anaerobic bacteria when grown under defined culture conditions

Incorporation of [methyl-3H]thymidine by obligate and facultative anaerobic bacteria when grown under defined culture conditions

Deoxyribonucleoside kinases in two aquatic bacteria with high specificity for thymidine and deoxyadenosine

Natural plasmid transformation in a high-frequency-of-transformation marine Vibrio strain

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