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1.
Hybrids between Escherichia coli K-12 and Salmonella typhosa which conserved a continuous K-12 chromosomal diploid segment extending from pro through ara to the strA locus were sensitive to plaque formation by wild-type λ. These partially diploid S. typhosa hybrids could be lysogenized with λ and subsequently induced to produce infectious phage particles. When the K-12 genes were segregated from a lysogenic S. typhosa hybrid, phage-productive ability was no longer detectable due to loss of a genetic region necessary for vegetative replication of λ. However, λ prophage was shown to persist in a quiescent state in the S. typhosa hybrid segregant with phage-productive ability being reactivated after replacement of the essential K-12 λ replication region. Low-frequency transduction and high-frequency transduction lysates containing the gal+ genes of S. typhosa were prepared by induction of λ-lysogenic S. typhosa hybrids indicating that the attλ site is chromosomally located in S. typhosa in close proximity to the gal locus as in E. coli K-12. After propagation in S. typhosa hybrids, λ was subject to restriction by E. coli K-12 recipients, thus establishing that S. typhosa does not perform the K-12 modification of λ deoxyribonucleic acid. Hybrids of S. typhosa, however, did not restrict λ grown previously on E. coli K-12. The K-12 genetic region required for λ phage production in S. typhosa was located within min 66 to min 72 on the genetic map of the E. coli chromosome. Transfer of an F-merogenote encompassing the 66 to 72 min E. coli chromosomal region to λ-insensitive S. typhosa hybrids enabled them to replicate wild-type λ. The λ-insensitive S. typhosa hybrid, WR4255, which blocks λ replication, can be mutagenized to yield mutant strains sensitive to λvir and λimm434. These WR4255 mutants remained insensitive to plaque formation by wild-type λ. 相似文献
2.
Carcinogenic N-nitrosamines were tested for their ability to induce λ 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. 相似文献
3.
The light-induced action of 8-methoxypsoralen (8-MOP) on λ phage and plasmids yields monoadducts and interstrand crosslinks. The survival and clear plaque mutation frequency in the phage photosensitized with 8-MOP and irradiated with UV at wavelength >320 nm are increased when the wild-type host (Escherichia coli uvr +) is subjected to UV irradiation (wavelength = 254 nm) prior to phage inoculation. These phenomena are known as “W reactivation” and “W mutagenesis.” It is shown that 8-MOP monoadducts in λ DNA induce clear mutations in the phage inoculated to UV-irradiated excision repair mutants of E. coli only when the error-prone repair is performed by MucA 2 ′ B, but not PolV (UmuD 2 ′ C) polymerase. The efficiency of the SOS repair (W reactivation) of 8-MOP monoadducts in plasmid and λ phage DNA also only increases with the presence of pKM101 plasmid muc + in E. coli uvr ?. 相似文献
4.
Henry Drexler 《Journal of virology》1973,12(5):1072-1077
A 10- to 15-min derepression of a λ prophage in a Gal− recipient during early infection with a transducing lysate of coliphage T1am will cause an increase in the efficiency of transduction of Gal+. An increase in the efficiency of transduction occurs when the donor is either nonlysogenic or lysogenic for λ; the increase is blocked by rifampin or chloramphenicol. With strain R901 it has been shown that efficient transduction can be blocked by treatment with rifampin after all chloramphenicol-sensitive steps have occurred. 相似文献
5.
Properties of a UV-sensitive Neurospora strain defective in pyrimidine dimer excision 总被引:1,自引:0,他引:1
A L Schroeder 《Mutation research》1974,24(1):9-16
The UV-sensitive Neurospora strain uvs-2 is known to resemble the excision-defective uvr mutants of E. coli K12 in being both excision-defective and highly UV mutable. As shown in this report, the uvs-2 strain also resembles the uvr mutants in its ability to remain photoreactivable when held in the dark for 2 h between UV-irradiation and photoreactivating light exposure, and in its maintenance of the same spontaneous deletion rate as wild type strains.Unlike the E. coli uvr mutants, however, this strain is sensitive to ionizing radiation and shows an increase in survival when held for 2 h in distilled water before plating (liquid-holding recovery [LHR]). The strain is three times more sensitive to X-rays than the wild type strain. It is also sensitive to nitrosoguanidine (MNNG). Sensitivity to UV, X-rays and MNNG appears to be under the control of a single gene.These properties suggest that the repair defect in the Neurospora uvs-2 mutant is different from those of the uvr mutants of E. coli K12. 相似文献
6.
Bacterial Cell Division Regulation: Lysogenization of Conditional Cell Division lon− Mutants of Escherichia coli by Bacteriophage Lambda
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The lon− mutants of Escherichia coli grow apparently normally except that, after temporary periods of inhibition of deoxyribonucleic acid synthesis, septum formation is specifically inhibited. Under these conditions, long, multinucleate, nonseptate filaments result. The lon− mutation also creates a defect such that wild-type bacteriophage λ fails to lysogenize lon− mutants efficiently and consequently forms clear plaques on a lon− host. Two lines of evidence suggest that this failure probably results from interference with expression of the λcI gene, which codes for repressor, or with repressor action:-(i) when a lon− mutant was infected with a λcII, cIII, or c Y mutant, there was an additive effect between the lon− mutation and the λc mutations upon reduction of lysogenization frequency; and (ii) lon− mutants permitted the growth of the λcro− mutant under conditions in which the repressor was active. The isolation of λ mutants (λtp) which gained the ability to form turbid plaques on lon− cells is also reported. 相似文献
7.
Summary Ultraviolet light sensitive mutants ofE. coli defective at theuvrA,uvrB oruvrC locus showed increased sensitivity to the lethal effects of mitomycin C when compared with theuvr
+ parental strain. In addition, DNA breakdown after treatment of cells with either mitomycin C or with ultraviolet light was greater in the parental strain carrying the activeuvr
+ genes than inuvr mutants. Thus, injuries produced by either mitomycin C or by ultraviolet light may be repaired by the same molecular mechanism which has been proposed and which involves defect excision, single strand breakdown and reconstruction of the DNA.With 2 Figures in the Text 相似文献
8.
Deise Fonseca Felício Leonardo da Silva Vidal Roberto Silva Irineu Alvaro Costa Leitão Wanda Almeida von Kruger Constança de Paoli Britto Angélica Cardoso Janine Simas Cardoso Claudia Lage 《DNA Repair》2013,12(1):63-72
Cisplatin is currently used in tumor chemotherapy to induce the death of malignant cells through blockage of DNA replication. It is a commonly used chemotherapeutic agent binding mono- or bifunctionally to guanines in DNA. Escherichia coli K12 mutant strains deficient in nucleotide excision repair (NER) were submitted to increasing concentrations of cisplatin, and the results revealed that uvrA and uvrB mutants are sensitive to this agent, while uvrC and cho mutants remain as the wild type strain. The time required for both gene expression turn-off and return to normal weight DNA in wild-type E. coli was not accomplished even after 4 h post-treatment with cisplatin, while the same process takes place within 1.5 h after ultraviolet radiation (UV). Besides, a heavily damaging action of cisplatin can be seen not only by persistent nicks on genomic DNA, but also by NER gene expression exceeding manifold that seen after equivalent lethal doses of UV. Moreover, cisplatin caused an increase in uvrB gene expression from its putative upstream promoter P3 in an SOS-independent manner. 相似文献
9.
《Mutation Research/Genetic Toxicology》1980,77(3):197-208
The antineoplastic drug daunorubicin and 15 other anthracyclines were tested for their ability to induce prophage λ in Escherichia coli K12. Prophage λ induction by daunorubicin was obtained in excision-repair deficient uvr− bacteria at doses about 3-fold lower than in excision-repair proficient uvr+ cells; this suggests that some of the lesions produced in DNA by daunorubicin are subject to excision repair and may be adducts. Daunorubicin seems to be converted to active species capable of causing prophage inducing lesions in DNA by bacterial enzymes. The antineoplastic and prophage inducing potencies of the anthracyclines were compared in a blind test. These two parameters were correlated for two thirds of the compounds. Such a correlation supports the idea that the antineoplastic activity of the anthracyclines is a consequence of their capacity to damage DNA. 相似文献
10.
11.
Temperature-Sensitive Modification and Restriction Phenotypes of an Escherichia coli dnaD Mutant 总被引:1,自引:0,他引:1
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A mutant of Escherichia coli temperature-sensitive for deoxyribonucleic acid synthesis, dnaD, was found to have temperature-sensitive modification and restriction phenotypes. In contrast to the original observation by Carl (1970), the mutant could support the growth of λ phage at 41 C. However, the λ phages thus produced were able to form plaques with normal plating efficiency only on E. coli C, a restriction-less strain, but not on E. coli K. Since the λ phages produced in the mutant at 30 C could form plaques equally well on both E. coli strains, it was concluded that the dnaD mutant has a temperature-sensitive modification phenotype. Furthermore, since the dnaD mutant allowed some growth of unmodified λ·C phages at 41 C but less at 30 C, the mutant is also temperature sensitive in restriction. The relationship, if any, between temperature-sensitive deoxyribonucleic acid synthesis and temperature-sensitive modification-restriction in the dnaD mutant is not known. Similar experiments were done with three dnaC mutants and one dnaA mutant. Two dnaC mutants were found to have altered restriction phenotypes at 41 C, but none of the mutants were defective in modification. 相似文献
12.
The effect of glutathione (GSH) on the ultraviolet (UV) induction of lambda prophage was investigated in lysogenic Escherichia coli. The data showed that extracellular GSH could inhibit the UV induction of lambda prophage. The inhibitory rates were concentration
dependent, and the maximal rate obtained was 94% with 3.0 M GSH. The effect was also measured in three different lambda lysogens:
a wild-type strain (wt), an isogenic GSH-deficient strain, and an isogenic strain producing increased amounts of GSH. The
result showed that when subjected to UV irradiation (254 nm, 60 J m−2), GSH-deficient strain was approximately fivefold more sensitive to be lysed than wt, whereas the strain with higher intracellular
GSH levels was only 28% susceptible to be lysed. With electron spin resonance and spin trapping techniques, we observed that
free radical signals occurred in the suspensions of UV irradiated lysogenic cells and the intensity of signals was influenced
by GSH levels. These results indicate that GSH can significantly inhibit the UV induction of lambda prophage, and that this
effect is correlated to its capacity to scavenge free radicals generated after UV irradiation. 相似文献
13.
Some Properties of Excision-defective Recombination-deficient Mutants of Escherichia coli K-12 总被引:22,自引:16,他引:6
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Strains of Escherichia coli that carry the mutation uvrA6 show no measurable excision of pyrimidine dimers and are easily killed by ultraviolet (UV) light, whereas strains that carry recA13 are defective in genetic recombination and are also UV-sensitive. An Hfr strain carrying uvrA6 was crossed with an F− strain carrying recA13. Among the recombinants identified, one carrying uvrA recA proved to be of exceptional sensitivity to UV light. It is estimated from the UV dose (0.2 erg/mm2 at 253.7 nm) required to reduce the number of colony-forming cells by one natural logarithm that about 1.3 pyrimidine dimers were formed in a genome of 5 × 106 base pairs for each lethal event. This double mutant is 40 times more UV-sensitive than the excision-defective strain carrying uvrA6. The replication of one pyrimidine dimer is generally a lethal event in strains carrying recA13. Spontaneous breakdown and UV-induced breakdown of the deoxyribonucleic acid (DNA) of cells of the various genotypes were estimated by growing the cells in medium containing 3H-thymidine and measuring both acid-precipitable and acid-soluble radioactivity. The UV-induced degradation in strains with recA13 did not require the uvr+ genes and hence appears to depend upon a mechanism other than dimer excision. The greater level of survival after irradiation in Rec+ as compared to Rec− bacteria may be due to a recovery mechanism involving the reconstruction of the bacterial chromosome through genetic exchanges which occur between the newly replicated sister duplexes and which effectively circumvent the damaged bases remaining in the DNA. 相似文献
14.
Fine Structure Mapping, Complementation, and Physiology of ESCHERICHIA COLI hfl Mutants 总被引:11,自引:1,他引:10
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Six of seven hfl mutations of Escherichia coli K12, characterized by high frequencies of lysogenization by phage lambda and λcIII mutants, are shown to be tightly linked to, but not within, the purA locus. All six hfl mutations are recessive to wild type in hfl+/hfl merodiploids and all lie in a single complementation group, located just counterclockwise from the purA locus. All six mutations confer a slightly increased resistance to penicillin and rifamycin and a slightly increased sensitivity to sodium dodecyl sulfate. Some cases of intragenic complementation and intragenic recombination were observed. It is argued that the hfl+ gene determines the synthesis of a protein which antagonizes lysogenization by phage lambda. It is further argued that the function of the λcIII gene product is to negate the antagonistic effect of this hfl+ protein. 相似文献
15.
Complementation between uvr mutants of Streptococcus pyogenes 总被引:1,自引:0,他引:1
Horst Malke 《Molecular & general genetics : MGG》1970,109(3):278-284
Summary From the group A streptococcal strains K 56 and 56 188, respectively 13 and 6 nitrosoguanidine-induced uvr mutants were isolated and used in complementation experiments employing strain 56 188 and its derivatives as donors in phage A 25-mediated heterologous transductions. When A 25 propagated on wild type or on 2 of the six 56 188-derived uvr mutants was used to infect 4 of the uvr recipient strains, a substantial increase in survivors of UV irradiation was found over those observed in selfing experiments or control experiments without phage. Less than 1% of the UV survivors had stably integrated the uvr
+ allele. The remaining 4 uvr donors failed to complement the 4 above-mentioned recipients, indicating that the strains in question fell into 2 complementation groups.Nine of the 13 K 56-derived mutants, which in contrast to the others were characterized by non-reversibility to UV resistance, did not even show an increase in UV survivors when infected with phage grown on wild type. The possibility is discussed that these strains might carry second mutations affecting UV sensitivity which, however, did not appear to be of the rec type. 相似文献
16.
Chad W. Euler Barbara Juncosa Patricia A. Ryan Douglas R. Deutsch W. Michael McShan Vincent A. Fischetti 《PloS one》2016,11(1)
Streptococcus pyogenes is a human commensal and a bacterial pathogen responsible for a wide variety of human diseases differing in symptoms, severity, and tissue tropism. The completed genome sequences of >37 strains of S. pyogenes, representing diverse disease-causing serotypes, have been published. The greatest genetic variation among these strains is attributed to numerous integrated prophage and prophage-like elements, encoding several virulence factors. A comparison of isogenic strains, differing in prophage content, would reveal the effects of these elements on streptococcal pathogenesis. However, curing strains of prophage is often difficult and sometimes unattainable. We have applied a novel counter-selection approach to identify rare S. pyogenes mutants spontaneously cured of select prophage. To accomplish this, we first inserted a two-gene cassette containing a gene for kanamycin resistance (KanR) and the rpsL wild-type gene, responsible for dominant streptomycin sensitivity (SmS), into a targeted prophage on the chromosome of a streptomycin resistant (SmR) mutant of S. pyogenes strain SF370. We then applied antibiotic counter-selection for the re-establishment of the KanS/SmR phenotype to select for isolates cured of targeted prophage. This methodology allowed for the precise selection of spontaneous phage loss and restoration of the natural phage attB attachment sites for all four prophage-like elements in this S. pyogenes chromosome. Overall, 15 mutants were constructed that encompassed every permutation of phage knockout as well as a mutant strain, named CEM1ΔΦ, completely cured of all bacteriophage elements (a ~10% loss of the genome); the only reported S. pyogenes strain free of prophage-like elements. We compared CEM1ΔΦ to the WT strain by analyzing differences in secreted DNase activity, as well as lytic and lysogenic potential. These mutant strains should allow for the direct examination of bacteriophage relationships within S. pyogenes and further elucidate how the presence of prophage may affect overall streptococcal survival, pathogenicity, and evolution. 相似文献
17.
We have investigated the production of prophage deletions in heat-induced λ lysogens of E. coli K12. Our results are indicative of a direct action of the heat-induced prophage in producing deletions. The temperature of 40° used for the experiments may be critical to prove this effect. The phage function involved in deletion formation is not known. 相似文献
18.
Analysis of the Role of Recombination and Repair in Mutagenesis of ESCHERICHIA COLI by Uv Irradiation 总被引:14,自引:0,他引:14
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Multiple mutant strains have been tested for their mimicry of the UV-mutagenesis deficiency of a recA single mutant. Revertants to histidine prototrophy and clear plaque mutants of lambda were scored to determine capacity for UV-mutagenesis. Nearly normal capacity was shown by a uvr+ recB- recF - strain, which shows almost no recA-dependent recombination, by uvr- recB+ recF - strains, which show almost no recA-dependent repair and by a uvrA- recB- recF- strain, which shows neither recA-dependent recombination nor repair. Since the uvr mutants can be assumed to show additionally no excision repair, these results may mean that UV-mutagenesis occurs during processes other than recombination and repair. Alternative hypotheses are discussed. The slight difference in mutagenic capacity was traced to the recF single mutation, which blocks the production of unmixed bursts of clear-plaque lambda mutants. Since this accounts for only about 10% of the mutations leading to clear-plaque mutants, it is suggested that there is more than one UV-mutagenic process. 相似文献
19.
Marianne De Paepe Laurent Tournier Elisabeth Moncaut Olivier Son Philippe Langella Marie-Agnès Petit 《PLoS genetics》2016,12(2)
Temperate phages, the bacterial viruses able to enter in a dormant prophage state in bacterial genomes, are present in the majority of bacterial strains for which the genome sequence is available. Although these prophages are generally considered to increase their hosts’ fitness by bringing beneficial genes, studies demonstrating such effects in ecologically relevant environments are relatively limited to few bacterial species. Here, we investigated the impact of prophage carriage in the gastrointestinal tract of monoxenic mice. Combined with mathematical modelling, these experimental results provided a quantitative estimation of key parameters governing phage-bacteria interactions within this model ecosystem. We used wild-type and mutant strains of the best known host/phage pair, Escherichia coli and phage λ. Unexpectedly, λ prophage caused a significant fitness cost for its carrier, due to an induction rate 50-fold higher than in vitro, with 1 to 2% of the prophage being induced. However, when prophage carriers were in competition with isogenic phage susceptible bacteria, the prophage indirectly benefited its carrier by killing competitors: infection of susceptible bacteria led to phage lytic development in about 80% of cases. The remaining infected bacteria were lysogenized, resulting overall in the rapid lysogenization of the susceptible lineage. Moreover, our setup enabled to demonstrate that rare events of phage gene capture by homologous recombination occurred in the intestine of monoxenic mice. To our knowledge, this study constitutes the first quantitative characterization of temperate phage-bacteria interactions in a simplified gut environment. The high prophage induction rate detected reveals DNA damage-mediated SOS response in monoxenic mouse intestine. We propose that the mammalian gut, the most densely populated bacterial ecosystem on earth, might foster bacterial evolution through high temperate phage activity. 相似文献
20.
Summary The relative yield (N
m/N) of fluorescent mutants Ind- after the transformation of Bacillus subtilis cells by means of UV-irradiated DNA is much higher in an uvr
- recipient than in an uvr
+ strain, when compared at equal fluence, but practically identical at equal survival. Ind- mutations are induced by UV-irradiation of separated single strands of transforming DNA. The H-strand is much more sensitive to the mutagenic action of UV light. Preliminary irradiation of competent recipient cells by moderate UV fluences increases the survival of UV-or -irradiated transforming DNA (W-reactivation) and the frequency of Ind- mutations (W-mutagenesis). During transfection of B. subtilis cells by UV-irradiated prophage DNA isolated from lysogenic cells B. subtilis (Ø105 c
+) c-mutants of the phage are obtained in high yield only in conditions of W-mutagenesis, i.e. in UV-irradiated recipient cells. These data show that there is no substantial spontaneous induction of error-prone SOS-repair system in the competent cells of B. subtilis. 相似文献