Formation of prototrophs in mixtures of two auxotrophic mutants of Serratia marcescens HY by a transducing bacteriophage produced by some auxotrophs

Summary Prototrophs arising in mixtures of two auxotrophs of Serratia marcescens strain HY are caused by a filtrable agent produced by one of the partners. 3. donors of this filtrable agent, HY/thyl, HY/ade11, and HY/thr2, were found among 16 auxotrophs, strain HY/thyl producing the agent of the highest activity. The prototrophic wildtype HY is not a donor. The agent does not enhance the growth of the auxotrophic recipient bacteria on minimal medium, therefore the increase in prototophs is not due to more spontaneous mutations. Dilution experiments showed that the reaction of one agent particle with a recipient-cell can cause a prototroph and that the number of recipient cells is not the limiting factor of prototroph formation.When the donor of a filtrable agent contained (besides the agent-inducing thyl-auxotrophy) a second auxotrophy of the same type as the recipient the relative frequency of prototrophs formed was much lower than that one produced by the HY/thyl filtrate. This indicates an influence of pseudoallelism of the markers on prototroph formation as would be expected by transfer of genetic material.The filtrable agent is non-dialyzable, precipitates by ammonium sulfate and is resistant to unspecific phosphodiesterase. When the filtrate was centrifuged in a CsCl density-gradient (24 hours at 35000 rpm) a band occurred at a density of 1.497 g/cm³ which contained the activities for prototroph and plaque formation. It contained also much material with an UV-absorption spectrum typical of phages. Electron micrographs revealed this material to consist of phage particles with hexagonal heads of 50 m diameter and a very short tail. These particles were named y phage.One strain, AX, of Serratia marcescens (among 47 tested) gave small, turbid plaques with filtrates from the known donors but not from other auxotrophs or HY. The plaque titer of y phage on AX was about the same as the transduction to prototrophy of HY/leu27. Phage y is a general-transducing bacteriophage of Serratia marcescens HY since 13 auxotrophic markers of strain HY and 10 of strain AX could be transduced. The low e.o.p. of y phage produced by HY/thyl, may therefore not be due to restriction by the AX cells but may indicate that this y phage is defective.