改进嗜甲基甲基ophilus的大规模培养，用于13C/15N标记和核糖核苷酸的随机部分氘代。
Improved large scale culture of Methylophilus methylotrophus for 13C/15N labeling and random fractional deuteration of ribonucleotides.
作者信息
Batey R T, Cloutier N, Mao H, Williamson J R
机构信息
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139, USA.
出版信息
Nucleic Acids Res. 1996 Dec 1;24(23):4836-7. doi: 10.1093/nar/24.23.4836.
Abstract
Isotopic labeling of RNA with 13C and 15N has become a routine procedure in structural studies by NMR spectroscopy. The methodology in this paper describes the random fractional deuteration of RNA using the obligate methylotropic bacterium, Methylophilus methylotrophus. This bacterium was grown using a non-deuterated carbon source in 52:48 D20/H20 and we have shown that all protons in the ribonucleotides except for the ribose H1 become 52% randomly fractionally deuterated. Improved growth conditions for this organism are also described that yield higher cell densities in liquid culture, which is applicable for all labeling procedures.
摘要
用¹³C和¹⁵N对RNA进行同位素标记已成为核磁共振光谱结构研究中的常规操作。本文中的方法描述了使用专性甲基营养菌甲基ophilus methylotrophus对RNA进行随机部分氘代。该细菌在52:48的D₂O/H₂O中使用非氘代碳源生长，我们已经表明，核糖核苷酸中除核糖H1外的所有质子都有52%被随机部分氘代。还描述了该生物体的改良生长条件，其在液体培养中可产生更高的细胞密度，适用于所有标记程序。
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