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Science 21 March 1969:
Vol. 163. no. 3873, pp. 1343 - 1348
DOI: 10.1126/science.163.3873.1343
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Articles

Mapping of Deletions and Substitutions in Heteroduplex DNA Molecules of Bacteriophage Lambda by Electron Microscopy

Barbara C. Westmoreland 1, Waclaw Szybalski 1, and Hans Ris 1

1 Department of Zoology and McArdle Laboratory, University of Wisconsin, Madison 53706

Electron microscopy of heteroduplex DNA molecules, composed of one strand of Escherichia coli phage lgr+ DNA annealed to the complementary DNA strand of a lgr deletion or substitution mutant, permits visualization, as well as precise measurements and mapping, of the unpaired single-stranded regions of nonhomology in the otherwise double-stranded molecules. In the lgrb2 mutant, the central segment (13 percent) of the lgr+ DNA molecule is shown to be deleted. In the hybrid phages lgri434 and lgri21 a segment of the right arm of the lgr+ genome (5.5 or 7.6 to 9 percent) is replaced by the corresponding immunity regions of phage 434 (3.3 percent or phage 21 (4 percent) DNA. The b5 region in the lgrb5 mutant appears to be identical to the i21 segment. From these data it is possible to estimate the size and posiion of those lgr genes which are replaced by the i434 and i21 segments. The method permits preparing complete physical maps of viral genomes with a precision heretofore unattainable.


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