A Restriction Map of Cauliflower Mosaic Virus DNA (Strain PV 147). Mapping of the Cleavage Sites of HhaI, SacI, AvaI, PvuII, PstI, XbaI, EcoRI, BglII, HincII, HpaII and HindII + III

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Abstract
The virion‐extracted DNA (Mr 5 × 106) 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, BglII, 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 1 site for HhaI and SacI, 2 for AvaI and PvuII, 3 for PstI, 4 for XbaI, 5 for EcoRI, 6 for BglII 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.