The generalized transducing phage F116 has been used to prepare lysates from fast-and slow-growing cultures of Pseudomonas aeruginosa strain 1. These lysates have been used to transduce a number of auxotrophic markers to prototrophy and the ratios of the numbers of transductants obtained with each lysate have been determined. Since the markers are those which have been mapped by conjugation in previous studies it has been possible to compare the ratios obtained for each marker with the relative position of the marker on the chromosome map. If the assumption is made that there is only one circular chromosome in P. aeruginosa strain 1 it is possible to suggest a way in which two apparently unlinked segments might be joined together. It is also possible to suggest that the chromosome replicates sequentially in two directions from a fixed origin.

SUMMARYThe order of replication of a series of genes in Pseudomonas aeruginosa has been studied in synchronized cultures using a method based on the technique of sequential mutagenesis. This technique relies on the increased susceptibility of the replication point of the bacterial chromosome to mutagenesis by N-methyl-N′-nitro-N-nitrosoguanidine. The genes studied were those previously mapped by conjugation and whose order of replication had been studied by an investigation of gene frequencies in exponential populations. The results are consistent with the idea that there is two-way replication of the chromosome of P. aeruginosa starting at a point near trp-1 and arg-6. They also confirm that the two linkage groups which have been found by conjugation replicate at different times. If the assumption is made that there is only one chromosome in P. aeruginosa, the results can be used to show how the two linkage groups may possibly be joined together and the order is such that there would have to be two sites of attachment for the sex factor FP2.