Predominant integration end products of infecting bacteriophage Mu DNA are simple insertions with no preference for integration of either Mu DNA strand

Abstract

The integration of 32P-labeled infecting Mu DNA into the Escherichia coli chromosome was investigated. Cleavage of the integrated Mu DNA with restriction endonuclease EcoRI, which cuts twice in the Mu genome, liberated the internal EcoRI fragment but not the left and right end fragments. The ends of the Mu genome became fused with host DNA at a variety of locations generating a smear of radioactive DNA fragments following cleavage with EcoRI. The predominant integration end products of infecting Mu DNA molecules are therefore generated by a mechanism which results in simple insertions and not cointegrates. Since predominantly simple insertions are found after infection (during lysogenization or lytic growth) but not after prophage induction, the transposition mode which is utilized appears to be a function of the source of the transposing DNA. Use of the integrated, 32P-labeled Mu DNA as a hybridization probe with the separated strands of Mu DNA or λ phages carrying various regions of Mu showed no strand preference in the integration process. Both labeled DNA strands at both ends of the Mu genome were integrated. These results suggest the lack of a site-specific recombination site in the genome; the simple insertions which are the end products of Mu DNA integration following infection appear to be generated by a separate pathway rather than by the resolution of cointegrate structures.