Growth of wildtype and mutant E. coli strains in minimal media for optimal production of nucleic acids for preparing labeled nucleotides.
Growth of wildtype and mutant E. coli strains in minimal media for optimal production of nucleic acids for preparing labeled nucleotides.
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野生型和突变型大肠杆菌菌株在基本培养基中的生长，以优化核酸生产，以制备标记的核苷酸。
DOI：
10.1007/s00253-010-2813-y
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发表时间：
2010
影响因子：
5
通讯作者：
Dayie,TKwaku
中科院分区：
文献类型：
--
作者：
Thakur,ChandarS;Brown,MargaretE;Sama,JacobN;Jackson,MelanthaE;Dayie,TKwaku
由于RNA位于大多数细胞过程的中心，因此需要合成大量由稳定同位素标记的核苷酸制成的RNA，以通过核磁共振（NMR）光谱来推进其结构和动力学的研究。获得标记核苷酸的一种特别有效的方法是从在补充有15 NH 4Cl和各种碳源的限定基本培养基中生长的细菌中收获这些核苷酸。考虑到NMR研究标记核酸所需的碳前体的高成本，评估常用菌株在标准基本培养基条件下的最佳生长变得重要。这方面的资料缺乏。在这项研究中，我们描述了生长forEscherichiacoli菌株K12，K10 zwf，和DL 323在三个基本培养基与同位素标记的碳源乙酸，甘油，甘油与甲酸盐相结合。在这三种培养基中，LeMaster-Richards和Studier培养基优于常用的M9培养基，并且都支持E.大肠杆菌生产核苷酸。然而，所有三个E的增长。与在甘油中的生长相比，在乙酸盐中的大肠杆菌减少了几乎两倍。对代谢途径的分析和先前的基因阵列研究有助于解释甘油和乙酸盐中的这种差异生长。这些研究将有助于选择性13 C-15 N同位素标记的核苷酸合成生物学上重要的RNA。
Since RNAs lie at the center of most cellular processes, there is a need for synthesizing large amounts of RNAs made from stable isotope-labeled nucleotides to advance the study of their structure and dynamics by nuclear magnetic resonance (NMR) spectroscopy. A particularly effective means of obtaining labeled nucleotides is to harvest these nucleotides from bacteria grown in defined minimal media supplemented with15NH4Cl and various carbon sources. Given the high cost of carbon precursors required for labeling nucleic acids for NMR studies, it becomes important to evaluate the optimal growth for commonly used strains under standard minimal media conditions. Such information is lacking. In this study, we characterize the growth forEscherichia colistrains K12, K10zwf, and DL323 in three minimal media with isotopic-labeled carbon sources of acetate, glycerol, and glycerol combined with formate. Of the three media, the LeMaster-Richards and the Studier media outperform the commonly used M9 media and both support optimal growth ofE. colifor the production of nucleotides. However, the growth of all threeE. colistrains in acetate is reduced almost twofold compared to growth in glycerol. Analysis of the metabolic pathway and previous gene array studies help to explain this differential growth in glycerol and acetate. These studies should benefit efforts to make selective13C-15N isotopic-labeled nucleotides for synthesizing biologically important RNAs.