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首页> 外文期刊>Applied and Environmental Microbiology >Estimating Bacterial Production in Marine Waters from the Simultaneous Incorporation of Thymidine and Leucine
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Estimating Bacterial Production in Marine Waters from the Simultaneous Incorporation of Thymidine and Leucine

机译:估计胸腺嘧啶和亮氨酸同时掺入在海水中的细菌产量

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We examined the simultaneous incorporation of [3H]thymidine and [14C]leucine to obtain two independent indices of bacterial production (DNA and protein syntheses) in a single incubation. Incorporation rates of leucine estimated by the dual-label method were generally higher than those obtained by the single-label method, but the differences were small (dual/single = 1.1 ± 0.2 [mean ± standard deviation]) and were probably due to the presence of labeled leucyl-tRNA in the cold trichloroacetic acid-insoluble fraction. There were no significant differences in thymidine incorporation between dual- and single-label incubations (dual/ single = 1.03 ± 0.13). Addition of the two substrates in relatively large amounts (25 nM) did not apparently increase bacterial activity during short incubations (<5 h). With the dual-label method we found that thymidine and leucine incorporation rates covaried over depth profiles of the Chesapeake Bay. Estimates of bacterial production based on thymidine and leucine differed by less than 25%. Although the need for appropriate conversion factors has not been eliminated, the dual-label approach can be used to examine the variation in bacterial production while ensuring that the observed variation in incorporation rates is due to real changes in bacterial production rather than changes in conversion factors or introduction of other artifacts.
机译:我们检查了[3H]胸苷和[14C]亮氨酸的同时掺入,以在一次孵育中获得两个独立的细菌生产指数(DNA和蛋白质合成)。双标记法估计的亮氨酸掺入率通常高于单标记法,但差异很小(双/单= 1.1±0.2 [平均值±标准差]),可能是由于冷的三氯乙酸不溶级分中存在标记的亮氨酰tRNA。双标记和单标记孵育之间的胸腺嘧啶核苷掺入没有显着差异(双/单= 1.03±0.13)。在短时间孵育(<5小时)期间,以相对较大的量(25 nM)添加两种底物显然不会增加细菌活性。通过双标记方法,我们发现在切萨皮克湾的深度剖面上,胸苷和亮氨酸的掺入率存在协变量。基于胸苷和亮氨酸的细菌产量估算值相差不到25%。尽管没有消除对适当转化因子的需求,但双标记法可用于检查细菌产生的变化,同时确保观察到的掺入率变化是由于细菌产生的实际变化而不是转化因子的变化引起的或介绍其他文物。

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