Single-cell RNA content of natural marine planktonic bacteria measured by hybridization with multiple 16S rRNA-targeted fluorescent probes
Single-cell RNA content of natural marine planktonic bacteria measured by hybridization with multiple 16S rRNA-targeted fluorescent probes
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通过与多个 16S rRNA 靶向荧光探针杂交测量天然海洋浮游细菌的单细胞 RNA 含量
DOI：
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发表时间：
1994
期刊：
影响因子：
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通讯作者：
P. Kemp
中科院分区：
文献类型：
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作者：
Sanghoon Lee;P. Kemp
尽管细胞 RNA 有潜力作为研究自然环境中细菌活动的工具，但由于方法学的限制，细胞 RNA 很少被测量。在之前的一项研究中，我们使用多个 16S rRNA 靶向荧光探针定量分析了单细胞 RNA 含量。在本研究中，我们利用 6 个月期间（冬末至夏季）从沿海水域收集的 12 个天然浮游细菌样本来探索这种方法的潜力。我们通过溴化乙锭 (EtBr) 荧光分析测量了与多个探针、细胞 DNA 和 RNA 杂交的单细胞的荧光，以及 t3H]胸苷 (TdR) 掺入率。探针杂交细胞的荧光与 EtBr 荧光测定法测定的细胞 RNA 高度相关。冬季和夏季样本显示出不同的荧光频率分布模式。温度对TdR掺入率有很大的正向影响，但与细胞RNA含量呈负相关；只有在夏季高温下采集的样本显示 TdR 掺入与细胞 RNA 含量之间呈正相关。我们估计未用五个探针明显标记的细胞的 RNA 含量约为 0.3 fg。浮游细菌在水生生物中的作用
Despite the potential of cellular RNA as a tool for the study of bacterial activities in natural environments, cellular RNA has rarely been measured because of methodological limitations. In a previous study, we quantitatively analyzed single-cell RNA content with multiple 16S rRNA-targeted fluorescent probes. In the present study, we explore the potential of this approach with 12 samples of natural planktonic bacteria collcctcd from coastal water over a 6-month period (late winter to summer). We measured fluorescence from single cells hybridized with multiple probes, cellular DNA and RNA by ethidium bromide (EtBr) lluoromctry, and t3H]thymidine (TdR) incorporation rates. Fluorescence of the probehybridized cells was highly correlated to cellular RNA determined by EtBr fluorometry. Winter and summer samples showed different patterns of fluorescence-frequency distribution. Temperature had a large positive effect on TdR incorporation rates, but there was a negative correlation with cellular RNA content; only the samples taken at elevated summer temperatures showed a positive relationship between TdR incornoration and cellular RNA content. We estimated that the RNA content of the cells not visibly labeled wiih five probes was ~0.3 fg. The role of bacterioplankton in the aquatic
影响因子：
56.9
作者：
DELONG, EF;WICKHAM, GS;PACE, NR
通讯作者：
PACE, NR