Shapovalova N I, Zheleznaia L A, Matvienko N N, Matvienko N I
Biokhimiia. 1994 Apr;59(4):485-93.
The new site-specific endonuclease BspKT5I free from interfering impurities has been isolated from thermophilic soil bacteria Bacillus species KT5 by successive chromatography on blue agarose, hydroxyapatite and heparin-Sepharose. BspKT5I on double-stranded DNA recognizes the sequence 5'-CTGAAG16N decreases 3'-GACTTC14N increases and cleaves the DNA at the recognition site as indicated by the arrows to form dinucleotide 3'-protruding termini. The isolated endonuclease is an isoschisomer of Eco57I. However, unlike Eco57I, it is not stimulated by S-adenosylmethionine (SAM) and can therefore be related to subclass IIs but not to IV, as Eco57I. In addition, endonuclease BspKT5I, unlike Eco57I, has no methylase activity.
通过在蓝色琼脂糖、羟基磷灰石和肝素-琼脂糖上连续层析，从嗜热土壤细菌芽孢杆菌属KT5中分离出了不含干扰杂质的新型位点特异性内切核酸酶BspKT5I。BspKT5I在双链DNA上识别序列5'-CTGAAG16N减少3'-GACTTC14N增加，并在识别位点如箭头所示切割DNA，形成二核苷酸3'-突出末端。分离出的内切核酸酶是Eco57I的同裂酶。然而，与Eco57I不同，它不受S-腺苷甲硫氨酸（SAM）刺激，因此可能与IIs亚类相关，而不像Eco57I与IV亚类相关。此外，内切核酸酶BspKT5I与Eco57I不同，没有甲基化酶活性。
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