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[大肠杆菌CK细胞中DNA甲基化酶的鉴定]

[Idnetification of DNA methylases in Escherichia coli CK cells].

作者信息

Nikol'skaia I I, Vanioshin B F

出版信息

Biokhimiia. 1975 Sep-Oct;40(5):1081-6.

PMID:764884
Abstract

E. coli CK cell are found to contain metylase which catalyses the incorporation of CH33-groups into tissue and phage DNAs in vitro in the presence of S-adenosyl-L-methionine, the donor of methyl groups. The enzyme was precipitated by (NH4)2SO4 of 30-60% saturation, which increased its specific activity in 1.9 times. Metylase was active both in phosphate and Tris. HCl buffers, pH 6.5-7.5 and did not require Mg2+, EDTA and dithiotreithol. The enzyme recognises certain nucleotide sequences in all the DNAs studied and has more wide specificity as compared with the enzyme from E. coli B. Methylase from E. coli CK developed the highest activity with thymus DNA. Methylase from rat liver nuclei turned to be inactive with bacteriophage DNA.

摘要

发现大肠杆菌CK细胞含有甲基化酶,该酶在甲基供体S-腺苷-L-甲硫氨酸存在的情况下,能在体外催化将CH₃基团掺入组织和噬菌体DNA中。该酶在30%-60%饱和度的硫酸铵中沉淀,其比活性提高了1.9倍。甲基化酶在磷酸盐和Tris.HCl缓冲液(pH 6.5-7.5)中均有活性,并且不需要Mg²⁺、EDTA和二硫苏糖醇。该酶能识别所有研究的DNA中的特定核苷酸序列,与来自大肠杆菌B的酶相比,具有更广泛的特异性。来自大肠杆菌CK的甲基化酶对胸腺DNA活性最高。来自大鼠肝细胞核的甲基化酶对噬菌体DNA无活性。

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