Conversion of Vibrio eltor MAK757 to classical biotype: role of phage PS166.
(49/1542)
Temperate phage PS166 infection of Vibrio eltor MAK757 resulted in complete changes in all biotype-specific determinants. About 10% of the PS166 lysogens of MAK757 lost their eltor-specific determinants, namely, the ability to produce soluble hemolysin, cell-associated hemagglutinin for chicken erythrocytes, and resistance to polymyxin B, as well as resistance to Mukherjee's group IV phage and sensitivity to eltor phage e4. These lysogens were found to have acquired the properties of classical strains, most significantly becoming sensitive to group IV phage but resistant to eltor-specific e4. The remainder of these lysogens, however, retained their parental biotype and serotype but acquired auxotrophy for glycine and histidine. The differential behavior of the two types of lysogen was due to the integration of the phage PS166 genome at different locations in the host chromosome. A 800-bp BglII fragment was found to contain the attP site. Phage PS166 has a polyhedral head (95 nm in diameter) and a contractile tail (98 nm in length). The phage chromosome is a linear double-stranded DNA of 110 kb and a G + C content of 58.7%. (+info)
Mechanism of phage PS166-mediated biotype conversion in Vibrio cholerae: role of the hlyA locus.
(50/1542)
Temperate phage PS166 lysogens of Vibrio eltor MAK757 biotype eltor belong to two major categories. Seventy percent of the lysogens acquire auxotrophy for glycine and histidine and maintain their parental biotype. About 10% of the lysogens become Cys(-) or Cys(-) Met(-) and are converted to the classical biotype with complete changes in all biotype-specific determinants. PCR and RFLP analysis revealed that in the latter lysogens, the phage genome integrated at the hlyA locus, whereas the same locus remained unaffected in lysogens that retained their parental biotype. These results suggest that the two types of lysogens arose due to integration of the phage genome at two different locations on the chromosome. A restriction map of the phage genome was constructed using AvaII and BglII. An 800-bp BglII fragment carrying the attP site, located at one of the termini of the phage genome, was used to distinguish the two classes of lysogen. (+info)
mut-25, a mutation to mutator linked to purA in Escherichia coli.
(51/1542)
The mutation mut-25 that results in a mutator phenotype is closely linked to purA on the chromosome of Escherichia coli. The gene order in this region is ampA mut-25 purA. purA mut-25 double mutants retained mutator activity indicating that mut-25 is not a mutation in the purA gene. The repair mutations uvrA6, recA56, and exrA1 had no effect on mutation frequencies in mut-25 strains, and mut-25 strains were normally resistant to ultraviolet irradiation. Frequencies of host range mutations were not increased in phages T1, T2, and T7 grown on mut-25 strains. mut-25 could act trans, reverting the trpA46 mutation either on the chromosome or on an F episome. The transitions AT yields GC (adenine-thymine yields guanine-cytosine) and GC yields AT were induced by mut-25. (+info)
Growth of bacteriophages MS2 and T7 on streptomycin-resistant mutants of Escherichia coli.
(52/1542)
Streptomycin-resistant mutants of an Hfr strain of Escherichia coli K were examined for their ability to support the growth of male-specific ribonucleic acid phage MS2 and female-specific deoxyribonucleic acid phage T7. Normally, the Hfr strain allows propagation of MS2 and is lysed by it (efficiency of plating equal to 1), whereas the same strain restricts propagation of T7 and is not lysed by it (efficiency of plating smaller than 10-7). Twenty-four isolates out of 26 independently obtained streptomycin-resistant mutants are partially or completely derestricted for propagation of T7; efficiency of plating of T7 in such strains ranges from 10-3-1. Depending on their response to plating of MS2 and T7, the streptomycin-resistant mutants can be divided into four classes. The mutants in all four classes continue to be "male" in conjugation with F- strains. Genetic analysis is presented to show that restriction of MS2, derestriction of T7, and resistance to streptomycin are the pleiotropic effects of a single mutation at the strA locus. (+info)
Purification of closed circular lambda deoxyribonucleic acid and its sedimentation properties as a function of Sodium chloride concentration and ethidium binding.
(53/1542)
The sedimentation of circular lambda DNA suggests that the molecular undergoes significant changes in shape and super-coiling as the NaC1 concentration increases. Closed circular lambda DNA, species I, isolated and purified from superinfected immune bacteria, sediments in sucrose gradients of low ionic strength at a rate 2.0 times faster than linear lambda DNA, species III. The addition of ethidium causes the sedimentation rate of species I DNA to decrease until enough dye is bound to remove 121 supercoils per molecule. At this point, species I co-sediments with nicked and nonsupercoiled species II. Futher additions of ethidium cause the sedimentation rate to increase until the relative rate of species I is again at least twice that of species III. This classical behavior is altered when NaC1 is present in the buffer. In 1.0 M NaC1 the changes in S are complex. Initially, species I sediments 1.55 times faster than species III. Titration with ethidium caused a decrease in S to an early minimum value, than an increase to a first maximum, followed by a decrease to the S of species II. At this point enough dye has intercalated to remove 208 superhelical turns. Further additions of dye introduce supercoils and cause S to increase again. In 0.1 to 0.4 M NaC1 the relative S of species I is 1.69 and 1.59, respectively. If titrated with ethidium, S first increases to a maximum value then decreases to the minimum rate when enough dye is bound to remove 158 and 183 supercoils, respectively. The results indicate an increase in the superhelix density from 0.026 turns per 10 base pairs in buffer alone to 0.045 in the same buffer with 1.0 M NaC1. If this change in superhelix density results from a concomitant change in the average rotation angle between base pairs in the Watson-Crick helix, the addition of 1.0 M NaC1 alters the rotation angle by 0.68 degrees per base pair. (+info)
Host DNA degradation after infection of Escherichia coli with bacteriophage T4: dependence of the alternate pathway of degradation which occurs in the absence of both T4 endonuclease II and nuclear disruption on T4 endonuclease IV.
(54/1542)
Escherichia coli cells infected with T4 phage which are deficient in both nuclear disruption and endonuclease II exhibit a pathway of host DNA degradation which does not occur in cells infected with phage deficient only in endonuclease II. This alternate pathway of host DNA degradation requires T4 endonuclease IV. (+info)
Sunlight-induced propagation of the lysogenic phage encoding cholera toxin.
(55/1542)
In toxigenic Vibrio cholerae, the cholera enterotoxin (CT) is encoded by CTXPhi, a lysogenic bacteriophage. The propagation of this filamentous phage can result in the origination of new toxigenic strains. To understand the nature of possible environmental factors associated with the propagation of CTXPhi, we examined the effects of temperature, pH, salinity, and exposure to direct sunlight on the induction of the CTX prophage and studied the transmission of the phage to potential recipient strains. Exposure of cultures of CTXPhi lysogens to direct sunlight resulted in approximately 10,000-fold increases in phage titers. Variation in temperature, pH, or salinity of the culture did not have a substantial effect on the induction of the prophage, but these factors influenced the stability of CTXPhi particles. Exposure of mixed cultures of CTXPhi lysogens and potential recipient strains to sunlight significantly increased both the in vitro and in vivo (in rabbit ileal loops) transduction of the recipient strains by CTXPhi. Included in these transduction experiments were two environmental nontoxigenic (CTXPhi(-)) strains of V. cholerae O139. These two O139 strains were transduced at high efficiency by CTXPhi, and the phage genome integrated into the O139 host chromosome. The resulting CTXPhi lysogens produced biologically active CT both in vitro and in rabbit ileal loops. This finding suggests a possible mechanism explaining the origination of toxigenic V. cholerae O139 strains from nontoxigenic progenitors. This study indicates that sunlight is a significant inducer of the CTX prophage and suggests that sunlight-induced transmission of CTXPhi may constitute part of a natural mechanism for the origination of new toxigenic strains of V. cholerae. (+info)
Prophage induction of Escherichia coli (lambda) by N-nitrosamines.
(56/1542)
Carcinogenic N-nitrosamines were tested for their ability to induce lambda in a lysogenic strain of Escherichia coli K-12 (58-161 F+ Dimethylnitrosamine, di-n-propylnitrosamine, methyl-n-propylnitrosamine, and N-nitrosopiperidine were shown to be inducers of prophage. (+info)