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Detection of Horizontal Gene Transfer by Natural Transformation in Native and Introduced Species of Bacteria in Marine and Synthetic Sediments

机译:通过自然转化检测海洋和合成沉积物中原生和引进细菌的水平基因转移

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Both naturally occurring marine sediments and artificial sediments were used as supports for natural transformation of marine bacteria. While transformation of Pseudomonas stutzeri ZoBell suspended in artificial seawater was not detected when recipient cells and rifampin resistance DNA were loaded onto sterile sediment columns, transformation could be detected at frequencies 4 to 20 times that of spontaneous resistance when recipient cells and rifampin resistance DNA were loaded onto sterile sediment columns. Treatment of these columns with DNase I reduced transformation frequencies to levels comparable to those of spontaneous-resistance frequencies. Sediments with higher organic contents supported higher frequencies of transformation than did those with lower amounts of organic matter. Transformation was also detected when recipient cells and DNA were loaded on columns prepared from nonsterile sediments, although the frequencies of transformation were lower than when sterile sediments were used. Finally, nonsterilized sediments that were not supplemented with laboratory strains did not support detectable levels of transformation in sediment columns, but when these same sediments were transferred to filters and placed on complex media, transformation was detected at a frequency three times that for spontaneous resistance. This transformation frequency was partially reduced to levels near that for spontaneous resistance by the addition of DNase I to sediment filters. These results indicate that marine sediments facilitate the uptake and expression of exogenous DNA by transformable marine bacteria and that sediments are a more likely niche for natural transformation than the water column in the marine environment.
机译:天然存在的海洋沉积物和人工沉积物都被用作海洋细菌自然转化的载体。当将受体细胞和利福平抗性DNA加载到无菌沉淀柱上时,未检测到悬浮在人工海水中的斯氏假单胞菌ZoBell的转化,但当加载受体细胞和利福平抗性DNA时,自发抗性的频率为4至20倍到无菌沉淀柱上。用DNase I处理这些色谱柱可将转化频率降低到与自发抗性频率相当的水平。有机物含量较高的沉积物比有机物含量较低的沉积物支持更高的转化频率。当将受体细胞和DNA装载到由非无菌沉淀物制备的柱子上时,也检测到了转化,尽管转化的频率比使用无菌沉淀物时要低。最后,未添加实验室菌株的未灭菌沉积物不支持在沉积物柱中检测到可检测的转化水平,但是当将这些相同的沉积物转移至过滤器并置于复杂介质中时,检测到的转化率是自发抵抗力的三倍。通过向沉淀物过滤器中添加DNase I,该转化频率被部分降低至接近自发抗性的水平。这些结果表明,海洋沉积物促进了可转化海洋细菌对外源DNA的吸收和表达,并且与海洋环境中的水柱相比,沉积物更可能成为自然转化的利基。

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