Tandem pentuplication of a DNA segment in a derivative of bacteriophage P2: its use in the study of the mechanism of DNA annealing

From a tandem duplication mutant of phage P2, triplication, quadruplication and pentuplication forms were derived. They were recognized by decreased virion heat stability resulting from the increase in DNA content, and were confirmed by electron microscope heteroduplex mapping. These forms of partially repeated DNA are quite stable in P2 because of the low level of recombination typical of this phage. Under conditions normally employed for full DNA renaturation, these high order repeat chromosomes gave often incomplete renaturation over the repeated segments. Based on current models for DNA renaturation, several predictions were made and tested. The results, although not quantitatively exhaustive, indicated that base pairing proceeding from a nucleation site was sufficiently slow to allow a second nucleation to occur with a fair probability over a length of a few thousand base pairs.