Hughes J A, Brown L R, Ferro A J
J Bacteriol. 1987 Aug;169(8):3625-32. doi: 10.1128/jb.169.8.3625-3632.1987.
We have developed a new research tool for the study of S-adenosylmethionine (AdoMet) metabolism by cloning the coliphage T3 AdoMet hydrolase (AdoMetase; EC 3.3.1.2) gene into the M13mp8 expression vector. The recombinant bacteriophage clones expressed an AdoMetase activity in Escherichia coli like that found in T3-infected cells. High levels of AdoMetase expression impaired AdoMet-mediated activities such as dam and dcm methylase-directed DNA modifications and the synthesis of spermidine from putrescine. Expression vectors containing the cloned AdoMetase gene thus provide an alternate approach to the use of chemical inhibitors or mutants defective in AdoMet biosynthesis to probe the effect of AdoMet limitation.
我们通过将大肠杆菌噬菌体T3腺苷甲硫氨酸水解酶(腺苷甲硫氨酸酶;EC 3.3.1.2)基因克隆到M13mp8表达载体中,开发了一种用于研究S-腺苷甲硫氨酸(AdoMet)代谢的新研究工具。重组噬菌体克隆在大肠杆菌中表达出一种腺苷甲硫氨酸酶活性,类似于在T3感染细胞中发现的活性。高水平的腺苷甲硫氨酸酶表达损害了AdoMet介导的活性,如dam和dcm甲基化酶指导的DNA修饰以及由腐胺合成亚精胺。因此,含有克隆的腺苷甲硫氨酸酶基因的表达载体为使用化学抑制剂或AdoMet生物合成缺陷型突变体来探究AdoMet限制的影响提供了另一种方法。