Kur J, Koob M, Burkiewicz A, Szybalski W
McArdle Laboratory for Cancer Research, Madison, WI 53706.
Gene. 1992 Jan 2;110(1):1-7. doi: 10.1016/0378-1119(92)90437-t.
Integration host factor (IHF)-mediated protection against enzymatic methylation at ihf-overlapping sites provides the basis for this novel application of the Achilles' cleavage (AC) technique [Koob et al., Science 241 (1988) 1084-1086] for generating rare natural cleavage sites. When applying IHF-AC to plasmid, phage lambda, Escherichia coli and yeast genomes, only a few of the EcoRI, HinfI, and MboI sites (which overlapped the ihf sites) remained cleavable after prior methylation with the cognate M.EcoRI, M.HinfI, or Dam methyltransferases in the presence of IHF. Thus, IHF-AC essentially converted these enzymes into very rare cutters. The extent of cleavage could be controlled by varying the IHF:DNA ratio and temperature. Moreover, the method permits the genomic location and strength of the ihf sites to be determined.
整合宿主因子（IHF）介导的对ihf重叠位点酶促甲基化的保护作用，为阿喀琉斯切割（AC）技术[库布等人，《科学》241（1988）1084 - 1086]这一用于产生罕见天然切割位点的新应用提供了基础。当将IHF - AC应用于质粒、λ噬菌体、大肠杆菌和酵母基因组时，在用同源的M.EcoRI、M.HinfI或Dam甲基转移酶在IHF存在下进行预先甲基化后，只有少数EcoRI、HinfI和MboI位点（与ihf位点重叠）仍可被切割。因此，IHF - AC本质上把这些酶转化为了非常罕见的切割酶。切割程度可以通过改变IHF与DNA的比例和温度来控制。此外，该方法还能确定ihf位点在基因组中的位置和强度。
