Saint R B, Egan J B
Mol Gen Genet. 1979 Mar 9;171(1):79-89. doi: 10.1007/BF00274018.
The restriction enzymes BamHI, Bg/II, EcoRI, HindIII, PstI, XbaI and XhoI have been used to cleave DNA isolated from the related coliphages P2 and 186 for analysis on 1% agarose gels. Three approaches were used to map the sites of cleavage: a) analysis dependent upon the existence of cohesive termini and availability of viable P2-186 hybrids; b) analysis of double digests and redigests of isolated fragments with a second enzyme and c) analysis of partial digests by transfer to nitrocellulose and hybridization with a single fragment. This last approach and the results obtained from it are detailed in a separate paper (Saint and Egan, 1979). The number of sites of each enzyme are as follows: a) 186, BamHI-7, Bg/II-1, EcoRI-3, HindIII-2, PstI-22, XbaI-0 and Xho-I-1; b) P2, BamHI-3, Bg/II-2, EcoRI-3, HindIII-0, PstI-O, XbaI-1 and XhoI-O. All of these sites have been mapped with the exception of PstI for 186, where only the five sites in the right 35% (the control region) have been mapped.
限制性内切酶BamHI、Bg/II、EcoRI、HindIII、PstI、XbaI和XhoI已用于切割从相关的大肠杆菌噬菌体P2和186中分离的DNA,以便在1%琼脂糖凝胶上进行分析。采用了三种方法来绘制切割位点:a)基于粘性末端的存在和可行的P2 - 186杂种的可用性进行分析;b)用第二种酶对分离的片段进行双酶切和再酶切分析;c)通过转移到硝酸纤维素膜上并用单个片段进行杂交来分析部分酶切。最后一种方法及其获得的结果在另一篇论文中详细阐述(Saint和Egan,1979年)。每种酶的切割位点数如下:a)186,BamHI - 7个、Bg/II - 1个、EcoRI - 3个、HindIII - 2个、PstI - 22个、XbaI - 0个、Xho - I - 1个;b)P2,BamHI - 3个、Bg/II - 2个、EcoRI - 3个、HindIII - 0个、PstI - 0个、XbaI - 1个、XhoI - 0个。除了186的PstI切割位点外,所有这些位点都已绘制出来,其中186的PstI切割位点仅绘制了右侧35%(控制区域)的五个位点。