Waugh D S
Department of Physical Chemistry, Roche Research Center, Hoffmann-La Roche Inc., Nutley, NJ 07110, USA.
J Biomol NMR. 1996 Sep;8(2):184-92. doi: 10.1007/BF00211164.
A collection of genetic tools that can be used to manipulate amino acid metabolism in Escherichia coli is described. The set comprises 21 strains of bacteria, each containing a different genetic defect that is closely linked to a selectable transposon marker. These tools can be used to construct strains of E. coli with ideal genotypes for residue-specific, selective labeling of proteins with nearly any 15N-amino acid. By using strains which have been modified to contain the appropriate genetic lesions to control amino acid biosynthesis, dilution of the isotope by endogenous amino acid biosynthesis and scrambling of the label to other types of residues can be avoided.
本文描述了一组可用于操纵大肠杆菌氨基酸代谢的遗传工具。该组包括21种细菌菌株，每种菌株都含有与可选择转座子标记紧密连锁的不同遗传缺陷。这些工具可用于构建具有理想基因型的大肠杆菌菌株，以便用几乎任何15N氨基酸对蛋白质进行残基特异性、选择性标记。通过使用经过修饰以包含适当遗传损伤来控制氨基酸生物合成的菌株，可以避免内源性氨基酸生物合成对同位素的稀释以及标记向其他类型残基的混杂。
