Assignment of imino proton spectra of yeast phenylalanine transfer ribonucleic acid.
Assignment of imino proton spectra of yeast phenylalanine transfer ribonucleic acid.
复制标题
酵母苯丙氨酸转移核糖核酸的亚氨基质子谱的分配。
DOI：
10.1021/bi00275a010
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发表时间：
1983
期刊：
影响因子：
2.9
通讯作者：
Redfield,AG
中科院分区：
文献类型：
--
作者：
Roy,S;Redfield,AG
材料和方法酵母tRNAPhe购自Boehringer曼海姆，无需进一步纯化即可使用。未对样品进行分析，但我们先前已重复分析了此类样品，并发现活性在1.5 nmol受体/μ L范围内。通过将5 mg tRNA溶解于190 µ缓冲液（含20 mM EDTA、10 mM磷酸盐（Na+）和0.1 M NaCl，pH 7.0）中制备NMR样品。将其在微池中用100 mL相同的缓冲液透析两次，并用相同的但含有1 mM EDTA的缓冲液透析两次。最后的透析是针对100 mL相同的缓冲液进行的，当需要时，该缓冲液含有1 mM EDTA和5% D2 G用于锁定。根据需要，将MgCl 2加入到对零EDTA透析的样品中。在下文中，“高镁”是指已经向其中添加了15 mM MgCl 2的样品。所有光谱均在自制的270-MHz仪器上记录
Materials and MethodsYeast tRNAPhe was purchased from Boehringer Mannheim and used without further purification. Samples were not assayed, but we have previously repeatedly assayed such samples and found activities in the range of 1.5 nmol of ac-ceptance//!^. NMR samples were prepared by dissolving 5 mg of tRNA in 190 µ of buffercontaining 20 mM EDTA, 10 mM phosphate (Na+), and 0.1 M NaCl at pH 7.0. This was dialyzed twice in a microcell against 100 mL of the same buffer and twice against the same buffer but containing 1 mM EDTA. Final dialysis was against 100 mL of the same buffer containing, when desired, 1 mM EDTA and 5% D2G for lock. MgCl2 was added as required to samples that were dialyzed against zero EDTA. In whatfollows,“high magnesium” refers to samples to which 15 mM MgCl2 has been added. All spectra were recorded on a homemade 270-MHz instrument