Volovitch M, Drugeon G, Dumas J P, Haenni A L, Yot P
Eur J Biochem. 1979 Oct;100(1):245-55. doi: 10.1111/j.1432-1033.1979.tb02055.x.
The virion-extracted DNA (Mr5 x 10(6)) of cauliflower mosaic virus (CaMV) has three single-stranded interruptions. The mapping of this DNA using eleven restriction endonucleases (HhaI, SacI, AvaI, PvuII, PstI, XbaI, EcoRI, Bg/II, HincII, HpaII and HindII + III) is reported here. The existence of the three single-stranded breaks complicates the identification and the molecular weight determination of fragments produced by HpaII, HindIII and HindII + III. Indeed the electrophoretic mobility of some fragments in which a single-stranded discontinuity is located is modified, and the fluorescence of ethidium bromide complexed with these fragments is reduced as compared to that observed for the other fragments existing in a molar ratio. These drawbacks were overcome by performing experiments of nick-translation of CaMV DNA with Escherichia coli DNA polymerase I. FRom the data it follows that the CaMV DNA molecule bears bears 1 site for HhaI and SacI, 2 for AvaI and PvuII, 3 for PstI, 4 for XbaI, 5 for EcoRI, 6 for Bg/II and HincII, 11 for HpaII and 15 for HindII + III. The corresponding fragments have all been ordered and precisely located providing a suitable map for further investigations connected with the study of the fine structure and the function of the CaMV genome.
花椰菜花叶病毒（CaMV）的病毒粒子提取DNA（Mr5×10⁶）有三个单链中断处。本文报道了使用11种限制性内切酶（HhaI、SacI、AvaI、PvuII、PstI、XbaI、EcoRI、Bg/II、HincII、HpaII以及HindII + III）对该DNA进行的图谱绘制。这三个单链断裂的存在使由HpaII、HindIII和HindII + III产生的片段的鉴定和分子量测定变得复杂。实际上，一些存在单链间断的片段的电泳迁移率会发生改变，并且与以摩尔比存在的其他片段相比，与这些片段结合的溴化乙锭的荧光会降低。通过用大肠杆菌DNA聚合酶I对CaMV DNA进行缺口平移实验克服了这些缺点。从数据可知，CaMV DNA分子有1个HhaI和SacI位点、2个AvaI和PvuII位点、3个PstI位点、4个XbaI位点、5个EcoRI位点、6个Bg/II和HincII位点、11个HpaII位点以及15个HindII + III位点。相应的片段都已排序并精确定位，为与CaMV基因组精细结构和功能研究相关的进一步研究提供了合适的图谱。
