Mutants of P22 which have been located in the c2 repressor gene were examined. The most rightward "c2 mutation" was found to define a site that is necessary only for the establishment and not for the maintenance of repressor synthesis. We conclude that this site c27 is an analog of cy mutants in phage lambda which define a promotor for repression establishment (pre). The K5 mutation of P22 maps between c27 and all other c2 mutants. Examination of its biological behavior and direct measurement of repressor activity show that K5 does not affect c2 repression. A model to explain these findings implies that c27 and K5 affect transcripts of opposite directions. P22 c1 mutants do not allow c2 repressor synthesis and we conclude that the activity of c1 product (and presumably c3 product) at the site defined by c27 is necessary for repressor synthesis. The combined activity of c1 and c3 product at c27 is postulated to promote repressor synthesis and block transcription of vegatative phage genes to the right of K5. After repressor synthesis has been established, another site analogous to lambda prm is sufficient for repressor synthesis and c27 is no longer required. These observations and conclusions point to a very close analogy between repressor synthesis and control in phages P22 and lambda.

A temperature-sensitive simian virus 40 (SV40) mutant, tsTNG-1, has been isolated from nitrosoguanidine-treated and SV40-infected African green monkey kidney (CV-1) cultures. Replication of virus at the nonpermissive temperature (38.7 C) was 3,000-fold less than at the permissive temperature (33.5 C). Plaque formation by SV40tsTNG-1 deoxyribonucleic acid (DNA) on CV-1 monolayers occurred normally at 33.5 C but was grossly inhibited at 38.7 C. The time at which virus replication was blocked at 38.7 C was determined by temperature-shift experiments. In shift-up experiments, cultures infected for various times at 33.5 C were shifted to 38.7 C. In shift-down experiments, cultures infected for various times at 38.7 C were shifted to 33.5 C. All cultures were harvested at 96 hr postinfection (PI). No virus growth occurred when the shift-up occurred before 40 hr PI. Maximum virus yields were obtained at 96 hr PI when the shift-down occurred at 66 hr, but only about 15% of the maximum yield was obtained when the shift-down occurred at 76 hr PI. These results indicate that SV40tsTNG-1 contains a conditional lethal mutation in a late viral gene function. Mutant SV40tsTNG-1 synthesized T antigen, viral capsid antigens, and viral DNA, and induced thymidine kinase activity at either 33.5 or 38.7 C. The properties of the SV40 DNA synthesized in mutant-infected CV-1 cells at 33.5 or 38.7 C were very similar to those of SV40 DNA made in parental virus-infected cells, as determined by nitrocellulose column chromatography, cesium-chloride-ethidium bromide equilibrium centrifugation, and by velocity centrifugation in neutral sucrose gradients. Mutant SV40tsTNG-1 enhanced cellular DNA synthesis in primary cultures of mouse kidney cells at 33.5 and 38.7 C and also transformed mouse kidney cultures at 36.5 C. SV40tsTNG-1 was recovered from clonal lines of transformed cells after fusion with susceptible CV-1 cells and incubation of heterokaryons at 33.5 C, but not at 38.7 C. MATERIALS AND METHODS Cell lines. CV-1, an established line of African green monkey kidney cells, was grown in "R5a" medium containing 10% calf serum and 0.5% lactalbumin hydrolysate (13), or in Eagle's minimal essential 286

A slowly growing, polymyxin-sensitive mutant of Salmonella typhimurium was isolated. Wild-type phage P22 form plaques on the mutant at 5 x 10-4, the frequency observed on wild-type hosts. All P22 clear mutants form plaques with near normal frequency. The inability of the mutant to form plaques is correlated with an increase in lysogenization frequency. The cause of the increased lysogenization frequency is not known, but it is not the result of overproduction of cyclic adenosine 5'-monophosphate.