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1.
Evaluation of the Role of Syringomycin in Plant Pathogenesis by Using Tn5 Mutants of Pseudomonas syringae pv. syringae Defective in Syringomycin Production 总被引:7,自引:1,他引:6
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Syringomycin is a necrosis-inducing phytotoxin produced by Pseudomonas syringae pv. syringae. To determine whether syringomycin production is a determinant in virulence or pathogenicity, we isolated nontoxigenic (Tox−) Tn5-containing mutants and then quantitatively evaluated them for the ability to multiply and cause disease in immature sweet-cherry fruits. Transposon Tn5 was delivered to Tox+ strain B301D-R by using the suicide vector, pGS9, and the resultant kanamycin-resistant (Kmr) colonies were screened for changes in syringomycin production by testing for antibiosis against Geotrichum candidum. Southern blot analysis of KpnI-and EcoRI-digested DNA showed that 15 (0.3%) Tox− mutants were isolated which had Tn5 inserted into 1 of 14 distinct loci. Phenotypic characterization of the Tox− mutants identified three major groups, which were differentiated by pathogenicity and ability to cause a tobacco hypersensitive reaction (HR). The eight strains in group A were pathogenic (Path+) in cherry fruit assays, but the disease index was 17 to 66% lower (significant at P = 0.01) than for the parental Tox+ strain, B301D-R. The population dynamics of group A strains W4S770 and W4S116 in cherry fruits were, however, indistinguishable from that of strain B301D-R. The remaining seven Tox− strains were nonpathogenic; group B strain W4S2545 (Path− HR+) and group C strain W4S468 (Path− HR−) developed significantly lower populations (105 to 107 CFU per cherry fruit) 3 days after inoculation than strain B301D-R did (nearly 109 CFU per fruit). The data indicate that syringomycin is not essential for pathogenicity, but contributes significantly to virulence. 相似文献
2.
Differential Synthesis of Peritoxins and Precursors by Pathogenic Strains of the Fungus Periconia circinata
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Alice C. L. Churchill Larry D. Dunkle Walter Silbert Kevin J. Kennedy Vlado Macko 《Applied microbiology》2001,67(12):5721-5728
Pathogenic strains of the soilborne fungus Periconia circinata produce peritoxins with host-selective toxicity against susceptible genotypes of sorghum. The peritoxins are low-molecular-weight, hybrid molecules consisting of a peptide and a chlorinated polyketide. Culture fluids from pathogenic, toxin-producing (Tox+) and nonpathogenic, non-toxin-producing (Tox−) strains were analyzed directly by gradient high-performance liquid chromatography (HPLC) with photodiode array detection and HPLC-mass spectrometry to detect intermediates and final products of the biosynthetic pathway. This approach allowed us to compare the metabolite profiles of Tox+ and Tox− strains. Peritoxins A and B and the biologically inactive intermediates, N-3-(E-pentenyl)-glutaroyl-aspartate, circinatin, and 7-chlorocircinatin, were detected only in culture fluids of the Tox+ strains. The latter two compounds were produced consistently by Tox+ strains regardless of the amount of peritoxins produced under various culture conditions. In summary, none of the known peritoxin-related metabolites were detected in Tox− strains, which suggests that these strains may lack one or more functional genes required for peritoxin biosynthesis. 相似文献
3.
Cohen LW 《Journal of virology》1969,4(3):214-218
A mutant of bacteriophage P22 (Lys−) was isolated which shows a plaque morphology on mixed plates comparable to the r+ plaques of the T-even phages. When Lys− and normal Lys+ plaques are juxtaposed on a petri dish, the Lys+ plaque exhibits a flat side adjacent to the Lys− plaque. The mutant is identical to P22 under an electron microscope, is inactivated at the same rate by antiserum and heat, and has the same kinetics of attachment. It does not plate on Salmonella lysogenic for phage P22 nor on strain St/22. In liquid culture, the lysis of mutant infections in M9CAA medium is delayed between 20 and 40 min. Cells mixedly infected in M9CAA with Lys− and Lys+ phage lyse later than Lys+-infected cells and even later than Lys−-infected cells. In unsupplemented M9 medium, however, mixedly infected cells again lyse later than Lys+-infected cells, but Lys−-infected cells require more than 3 hr to lyse. In supplemented and unsupplemented M9 media, intracellular phage development and endolysin synthesis proceed in Lys− infections at least as rapidly as in Lys+-infected cells. In diluted infections, the latent and eclipse periods of Lys− and Lys+ infections are indistinguishable. The possible mechanisms involved in the control and timing of lysis are discussed. 相似文献
4.
Stefan Hertwig Andreas Popp Barbara Freytag Rudi Lurz Bernd Appel 《Applied microbiology》1999,65(9):3862-3866
To study phage-mediated gene transfer in Yersinia, the ability of Yersinia phages to transduce naturally occurring plasmids was investigated. The transduction experiments were performed with a temperate phage isolated from a pathogenic Yersinia enterocolitica strain and phage mixtures isolated from sewage. Small plasmids (4.3 and 5.8 kb) were transduced at a frequency of 10−5 to 10−7/PFU. However, we could not detect the transduction of any indigenous virulence plasmid (ca. 72 kb) in pathogenic Yersinia strains. Transductants obtained by infection with the temperate phage were lysogenic and harbored the phage genome in their chromosomes. 相似文献
5.
Conjugal Transfer of Bacteriophage Resistance Determinants on pTR2030 into Streptococcus cremoris Strains 总被引:16,自引:13,他引:3
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Agar surface conjugal matings were used to introduce heat-sensitive phage resistance (Hsp+) determinants carried on the conjugal plasmid pTR2030 into Streptococcus cremoris KH, HP, 924, and TDM1. Lactose-fermenting (Lac+) transconjugants were selected from matings of Lac− variants of S. cremoris KH, HP, 924, and TDM1 with Streptococcus lactis ME2 or a high-frequency donor, S. lactis T-EK1 (pTR1040, Lac+; pTR2030, Hsp+). For all of the S. cremoris strains examined, select Lac+ transconjugants were completely resistant to plaquing by their homologous lytic phages. In all cases the plaquing efficiencies were less than 10−9. Acquisition of a 30-megadalton plasmid (pTR2030) in the S. cremoris phage-resistant transconjugants was demonstrated by direct plasmid analysis, by hybridization with 32P-labeled probes, or by conjugal transfer of pTR2030 out of the phage-resistant transconjugants into a plasmid-cured recipient, S. lactis LM2302. Acid production, coagulation ability, and proteolytic activity of phage-resistant transconjugants in milk were comparable to those of their phage-sensitive parents. Further, S. cremoris phage-resistant transconjugants were not attacked by phage in starter culture activity tests, which included a 40°C incubation period. The results demonstrated that phage resistance determinants on pTR2030 could be conjugally transferred to a variety of S. cremoris strains and confer resistance to phage under conditions encountered during cheese manufacture. Phage-resistant transconjugants of S. cremoris M43 and HP were also constructed without the use of antiblotic markers to select conjugal recipients from mating mixtures. 相似文献
6.
Hilary K. DeBardeleben Elena S. Lysenko Ankur B. Dalia Jeffrey N. Weiser 《Journal of bacteriology》2014,196(14):2670-2680
The pathogenesis of the disease caused by Streptococcus pneumoniae begins with colonization of the upper respiratory tract. Temperate phages have been identified in the genomes of up to 70% of clinical isolates. How these phages affect the bacterial host during colonization is unknown. Here, we examined a clinical isolate that carries a novel prophage element, designated Spn1, which was detected in both integrated and episomal forms. Surprisingly, both lytic and lysogenic Spn1 genes were expressed under routine growth conditions. Using a mouse model of asymptomatic colonization, we demonstrate that the Spn1− strain outcompeted the Spn1+ strain >70-fold. To determine if Spn1 causes a fitness defect through a trans-acting factor, we constructed an Spn1+ mutant that does not become an episome or express phage genes. This mutant competed equally with the Spn1− strain, indicating that expression of phage genes or phage lytic activity is required to confer this fitness defect. In vitro, we demonstrate that the presence of Spn1 correlated with a defect in LytA-mediated autolysis. Furthermore, the Spn1+ strain displayed increased chain length and resistance to lysis by penicillin compared to the Spn− strain, indicating that Spn1 alters the cell wall physiology of its host strain. We posit that these changes in cell wall physiology allow for tolerance of phage gene products and are responsible for the relative defect of the Spn1+ strain during colonization. This study provides new insight into how bacteria and prophages interact and affect bacterial fitness in vivo. 相似文献
7.
Phage H, thought to be specific for Pasteurella pestis, was shown to plate efficiently on F− strains of Escherichia coli but not on F+, F′, or Hfr strains. The phage was adsorbed rapidly to F− strains but was not adsorbed to strains carrying F. Comparison with seven other reported female-specific phages showed that, although phage H was similar to the other phages in some characteristics, the exceptionally low efficiency of plating (<10−9) on F-containing cells makes phage H a particularly useful female-specific phage. 相似文献
8.
The marine phage ΦHSIC has been previously reported to enter into a lysogenic relationship with its host, HSIC, identified as Listonella pelagia. This phage produces a variety of plaques on its host, including turbid and haloed plaques, from which lysogens were previously isolated. These lysogens were unstable during long-term storage at −80° C and were lost. When HSIC was reinfected with phage ΦHSIC, pseudolysogen-like interactions between the phage and its host were observed. The cells (termed HSIC-2 or HSIC-2e) produced high viral titers (1011 ml−1) in the absence of inoculating phage and yet reached culture densities of nearly 109 ml−1. Prophages were not induced by mitomycin C or the polyaromatic hydrocarbon naphthalene in cells harboring such infections. However, such cells were homoimmune to superinfection. Colonies hybridized strongly with a gene probe from a 100-bp fragment of the ΦHSIC genome, while the host did not. Analysis of chromosomal DNA preparations suggested the presence of a chromosomally integrated prophage. Phage adsorption experiments suggested that HSIC-2 was adsorption impaired. Because of the chromosomal prophage integration and homoimmunity, we interpret these results to indicate that ΦHSIC establishes a lysogenic relationship with its host that involves an extremely high level of spontaneous induction. This could be caused by a weak repressor of phage production. Additionally, poor phage adsorption of HSIC-2 compared to the wild type probably helped maintain this pseudolysogen-like relationship. In many ways, pseudolysogenic phage-host interactions may provide a paradigm for phage-host interactions in the marine environment. 相似文献
9.
Specific Action of Sodium Dodecyl Sulfate on the Sex Factor of Escherichia coli K-12 Hfr Strains 总被引:13,自引:6,他引:7
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Noriko Inuzuka Sachiko Nakamura Manabu Inuzuka Munemitsu Tomoeda 《Journal of bacteriology》1969,100(2):827-835
A specific action of sodium dodecyl sulfate (SDS) on the sex (F) factor in the integrated state of Escherichia coli K-12 Hfr H strain is reported. Growth of Hfr cells in Penassay Broth containing SDS results in the elimination of part or all of the F factor, yielding low and nonfertile variants of defective Hfr type and F+ cells and also F− derivatives. Appearance of such variants was generally observed after the culture reached stationary phase. The frequencies of F− cells then increased. F− cells were usually isolated as the major population among survivors. Some defective variants of Hfr cells with an intermediate fertility between standard Hfr and F+ cells had lost sensitivity toward the male-specific ribonucleic acid phage M12. Other defective Hfr variants with as much or less fertility than standard F+ cells had also all lost sensitivity to phage M12. On single-colony isolation, they segregated nonfertile female H cells which, when infected with F, could restore high fertility with oriented transfer of the chromosome the same as that of the original Hfr H. Also, sensitivity to phage M12 was regained. Female H cells were characterized as those lacking fertility but still retaining a small segment of F or sfa locus at the original part of the chromosome, where newly infected F could attach. Similar results were obtained with two other Hfr strains. A possible mechanism of the specific action of SDS is discussed. 相似文献
10.
Plasmid DNA in Streptococcus cremoris Wg2: Influence of pH on Selection in Chemostats of a Variant Lacking a Protease Plasmid 总被引:21,自引:13,他引:8
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Cleared lysates of a proteolytic (Prt+) strain and a naturally occurring non-proteolytic (Prt−) variant of Streptococcus cremoris Wg2 contain equal amounts of covalently closed circular plasmid DNA. An analysis of this plasmid DNA by agarose gel electrophoresis revealed the presence of at least five different plasmid species in the Prt+ strain and only three plasmid species in the Prt− variant. Curing studies with acriflavine indicated that a 16-megadalton plasmid determined proteolytic activity in the Prt+ strain. In energy-limited chemostats inoculated with both strains it was observed that the Prt+ strain was replaced by the Prt− variant. This effect was most apparent when the pH of the culture was fixed at a value above 6.3. No selection for the Prt− variant was observed at pH 5.9. Since the two types of organisms contain equal amounts of plasmid DNA, it was concluded that the energy gain of the Prt− variants at pH values above 6.0 probably has to be found in protein synthesis rather than in plasmid DNA synthesis. 相似文献
11.
Evidence for a Plasmid-Linked Restriction-Modification System in Lactobacillus helveticus 总被引:2,自引:2,他引:0
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Clara G. de los Reyes-Gaviln Gaëtan K. Y. Limsowtin Laurent Schaud Monique Veaux Jean-Pierre Accolas 《Applied microbiology》1990,56(11):3412-3419
The presence of a restriction-modification (R/M) system against two bacteriophages, 328-B1 and hv, was demonstrated in three Lactobacillus helveticus strains, CNRZ 1094, CNRZ 1095, and CNRZ 1096. In addition, the burst size of phage 328-B1 in the three restrictive strains CNRZ 1094, CNRZ 1095, and CNRZ 1096 was reduced with respect to the values obtained in its propagating strain, CNRZ 328. Heating at 60°C did not inactivate the R/M system. Nonrestrictive variants from CNRZ 1094 were easily obtained under several culture conditions, but treatment with novobiocin at 42°C followed by storage at −20°C resulted in drastic elimination of the R+/M+ phenotype from all clones tested. Electrophoretic analysis of CNRZ 1094 nonrestrictive variants revealed the concomitant loss of a 34-kb plasmid. Four EcoRI fragments from the 34-kb plasmid were cloned in the Escherichia coli vector pACYC184. The use of one or several of these fragments as probes confirmed the plasmidic location of the genes responsible for the R/M system. These probes also showed the presence of R/M plasmids in the two other restrictive strains, CNRZ 1095 and CNRZ 1096. Lactose-fermenting ability and/or proteolytic capacity was not linked to the 34-kb plasmid. 相似文献
12.
Charlotte E. Egan Erin K. Daugherity Arlin B. Rogers Delbert S. Abi Abdallah Eric Y. Denkers Kirk J. Maurer 《PloS one》2013,8(6)
Non-alcoholic fatty liver disease (NAFLD) is a common disease with a spectrum of presentations. The current study utilized a lithogenic diet model of NAFLD. The diet was fed to mice that are either resistant (AKR) or susceptible (BALB/c and C57BL/6) to hepatitis followed by molecular and flow cytometric analysis. Following this, a similar approach was taken in congenic mice with specific mutations in immunological genes. The initial study identified a significant and profound increase in multiple ligands for the chemokine receptor CCR2 and an increase in CD44 expression in susceptible C57BL/6 (B6) but not resistant AKR mice. Ccr2−/− mice were completely protected from hepatitis and Cd44−/− mice were partially protected. Despite protection from inflammation, both strains displayed similar histological steatosis scores and significant increases in serum liver enzymes. CD45+CD44+ cells bound to hyaluronic acid (HA) in diet fed B6 mice but not Cd44−/− or Ccr2−/− mice. Ccr2−/− mice displayed a diminished HA binding phenotype most notably in monocytes, and CD8+ T-cells. In conclusion, this study demonstrates that absence of CCR2 completely and CD44 partially reduces hepatic leukocyte recruitment. These data also provide evidence that there are multiple redundant CCR2 ligands produced during hepatic lipid accumulation and describes the induction of a strong HA binding phenotype in response to LD feeding in some subsets of leukocytes from susceptible strains. 相似文献
13.
The integration of the lactose plasmid from lactic streptococci into the host chromosome could stabilize this trait for dairy fermentations. Sixty lactose-positive (Lac+) transductants of lactose- and proteinase-negative (Lac− Prt−) LM0220 were induced for temperature phage by UV irradiation or mitomycin C. Four of the transductants, designated KB18, KB21, KB54, and KB58, yielded lysates demonstrating less than one Lac+ transductant per 0.2 ml of phage lysate. Successive transferring in the presence of acriflavine did not yield Lac− segregants from KB18, KB21, KB54, or KB58, whereas Streptococcus lactis C2 (parent culture) and three other Lac+ transductants showed 12 to 88% conversion from Lac+ to Lac− within 6 to 10 repetitive transfers. When grown in continuous culture, KB21 did not show any Lac− variants in 168 h, while S. lactis C2 had 96% conversion from Lac+ to Lac− in 144 h. Agarose gel electrophoresis of plasmid DNA isolated from KB18, KB21, KB54, and KB58 revealed that the lactose plasmid, pLM2103, normally present in Lac+ transductants, was missing. This suggested integration of the transferred lactose plasmid into the chromosome. In contrast to phage lysates induced from S. lactis C2, which exhibited an exponential decrease in the number of Lac+ transductants after exposure to small doses of UV irradiation, the transduction frequency for lactose metabolism was stimulated by UV irradiation of lysates from KB58. The latter indicated chromosomal linkage for lac and that integration of the lactose genes plasmid into the chromosome had occurred. 相似文献
14.
Frequency of Bacteriocin Resistance Development and Associated Fitness Costs in Listeria monocytogenes 总被引:4,自引:0,他引:4
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A. Gravesen A.-M. Jydegaard Axelsen J. Mendes da Silva T. B. Hansen S. Knchel 《Applied microbiology》2002,68(2):756-764
Bacteriocin-producing starter cultures have been suggested as natural food preservatives; however, development of resistance in the target organism is a major concern. We investigated the development of resistance in Listeria monocytogenes to the two major bacteriocins pediocin PA-1 and nisin A, with a focus on the variations between strains and the influence of environmental conditions. While considerable strain-specific variations in the frequency of resistance development and associated fitness costs were observed, the influence of environmental stress seemed to be bacteriocin specific. Pediocin resistance frequencies were determined for 20 strains and were in most cases ca. 10−6. However, two strains with intermediate pediocin sensitivity had 100-fold-higher pediocin resistance frequencies. Nisin resistance frequencies (14 strains) were in the range of 10−7 to 10−2. Strains with intermediate nisin sensitivity were among those with the highest frequencies. Environmental stress in the form of low temperature (10°C), reduced pH (5.5), or the presence of NaCl (6.5%) did not influence the frequency of pediocin resistance development; in contrast, the nisin resistance frequency was considerably reduced (<5 × 10−8). Pediocin resistance in all spontaneous mutants was very stable, but the stability of nisin resistance varied. Pediocin-resistant mutants had fitness costs in the form of reduction down to 44% of the maximum specific growth rate of the wild-type strain. Nisin-resistant mutants had fewer and less-pronounced growth rate reductions. The fitness costs were not increased upon applying environmental stress (5°C, 6.5% NaCl, or pH 5.5), indicating that the bacteriocin-resistant mutants were not more stress sensitive than the wild-type strains. In a saveloy-type meat model at 5°C, however, the growth differences seemed to be negligible. The applicational perspectives of the results are discussed. 相似文献
15.
The Escherichia coli lon− mutants apparently are defective in the ability to recommence cell division after temporary periods of deoxyribonucleic acid (DNA) synthesis inhibition. They are also more susceptible to cell division inhibition by the basic dye, crystal violet (CV), than are lon+ strains. In enriched broth, the lon+ strain continued to grow and divide in the presence of CV, but lon− cell division was inhibited and filamentous growth resulted. In a supplemented minimal medium containing CV, lon− cell division was only temporarily inhibited. There was no detectable specific effect on DNA synthesis, although CV slowed the rate of mass increase in both media. Trichloroacetic acid-insoluble lipid synthesis was preferentially inhibited in both lon+ and lon− strains. In CV-containing enriched broth, diaminopimelic acid incorporation into trichloroacetic acid-insoluble compounds occurred at a rate greater than the rate of mass increase in both lon+ and lon− strains. In a CV-containing supplemented minimal medium, diaminopimelic acid was incorporated to a greater extent by lon− cells than by lon+ cells. 相似文献
16.
Characterization of Phage-Sensitive Mutants from a Phage-Insensitive Strain of Streptococcus lactis: Evidence for a Plasmid Determinant that Prevents Phage Adsorption 总被引:16,自引:13,他引:3
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A phage-insensitive strain of Streptococcus lactis, designated ME2, was used as a prototype strain for the study of mechanisms and genetics of phage resistance in the lactic streptococci. Mutants sensitive to a Streptococcus cremoris phage, ϕ18, were isolated at a level of 17% from cultures of ME2 after sequential transfer at 30°C. Phage-sensitive mutants of ME2 were not fully permissive to ϕ18. The efficiency of plating of ϕ18 on the mutants was 5 × 10−7 as compared with <10−9 for ϕ18 on ME2. Further characterization of the mutants showed that they efficiently adsorbed ϕ18 at levels of >99.8%, whereas ME2 adsorbed only 20 to 40% of ϕ18. These results suggest that increased phage susceptibility of the mutants may result from the loss of a mechanism that inhibits phage adsorption. Moreover, the high frequency of spontaneous mutation in ME2 indicates the involvement of an unstable genetic determinant in this phage defense mechanism. ME2 was shown to possess 13 plasmids ranging in size from 1.6 to 34 megadaltons. Of 40 mutants examined that had increased efficiencies of plating, all were missing a 30-megadalton plasmid, pME0030. These data suggest that pME0030 codes for a function that prevents phage adsorption. Further phenotypic characterization of the phage-sensitive mutants showed that some mutants were deficient in the ability to ferment lactose (Lac) and hydrolyze milk proteins (Prt). However, the Lac+ and Prt+ phenotype segregated independently of the phage-sensitivity phenotype. One phage-sensitive adsorption mutant, designated N1, was tested for susceptibility to 14 different phages. N1 showed increased capacity to adsorb 4 and to replicate 2 of these 14 phages, thereby indicating a phage resistance mechanism in ME2 that generalizes to phage interactions other than the specific ϕ18-ME2 phage-host interaction. These data provide evidence for a unique plasmid-linked phage defense mechanism in phage-insensitive strains of lactic streptococci. 相似文献
17.
Transduction of Methicillin Resistance in Staphylococcus aureus Dependent on an Unusual Specificity of the Recipient Strain 总被引:26,自引:11,他引:15
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Resistance to methicillin was transduced by phage 80 or 53 from two naturally occurring methicillin-resistant strains of Staphylococcus aureus to methicillin-susceptible recipient strains at frequencies of 10−7 to 10−9. Ultraviolet irradiation of transducing phage and posttransductional incubation at 30 C were essential for useful frequencies of transduction. Effectiveness as a recipient for this transduction was highly specific. Strain NCTC 8325 (PS47) in its native state was an ineffective recipient but became effective after it had received by transduction one of several penicillinase plasmids. This acquired effectiveness was retained in most cases after elimination of the plasmid by ethidium bromide treatment. Like the donor strain, the progeny were heterogeneous in the degree of their resistance to methicillin, which was expressed by a higher proportion of cells as the temperature of incubation was lowered from 37 to 30 C. Separate transductants varied widely in the degree of resistance acquired by transduction. Methicillin resistance was stable in the donor and transductant strains. We favored the interpretation that methicillin resistance in our strains was determined by a single chromosomal gene, although the possibility that it was determined by two or more closely linked genes could not be excluded. 相似文献
18.
Phage Resistance in a Phage-Insensitive Strain of Streptococcus lactis: Temperature-Dependent Phage Development and Host-Controlled Phage Replication 总被引:6,自引:5,他引:1
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Streptococcus lactis ME2 is a dairy starter strain that is insensitive to a variety of phage, including 18. The efficiency of plating of 18 on ME2 and N1 could be increased from <1 × 10−9 to 5.0 × 10−2 and from 7.6 × 10−7 to 2.1 × 10−2, respectively, when the host strains were subcultured at 40°C before plating the phage and the phage assay plates were incubated at 40°C. Host-dependent replication was demonstrated in N1 at 30°C and in N1 and ME2 at 40°C, suggesting the operation of a temperature-sensitive restriction and modification system in ME2 and N1. The increased sensitivity of ME2 and N1 to 18 at 40°C was also demonstrated by lysis of broth cultures and increased plaque size. ME2 grown at 40°C showed an increased ability to adsorb 18, indicating a second target for temperature-dependent phage sensitivity in ME2. Challenge of N1 with a 18 preparation that had been previously modified for growth on N1 indicated that at 40°C phage development was characterized by a shorter latent period and larger burst size than at 30°C. The evidence presented suggests that the high degree of phage insensitivity expressed by ME2 consists of a variety of temperature-sensitive mechanisms, including (i) the prevention of phage adsorption, (ii) host-controlled restriction of phage, and (iii) suppression of phage development. At 30°C these factors appear to act cooperatively to prevent the successful emergence of lytic phage active against S. lactis ME2. 相似文献
19.
Yi Sui Rujian Zhu Wei Hu Wei Zhang Hongbo Zhu Min Gong Lili Gao Ting Cao Tao Tang Bo Yu Tao Yang 《Translational oncology》2021,14(3)
Patients with prostate cancer (PCa) will eventually progress to castrate-resistant prostate cancer (CRPC) after androgen deprivation therapy (ADT) treatment. Prostate-specific antigen (PSA)−/lo cells which harbor self-renewing long-term tumor-propagating cells that can be enriched using ALDH+CD44+α2β1+ and can initiate tumor development may represent a critical source of CRPC cells. Our purpose was to find a peptide that specifically targets PSA−/lo PCa cells to retard the development of CRPC. PSA+ and PSA−/lo cells were successfully separated from LNCaP xenograft tumors after prostate- PSAP-GFP vector infection and FACS. A variety of PSA−/lo cells specifically targeting peptide (named as “TAP1” targeted affinity peptide 1) was identified by using phage display library screening. The highest binding rate in TAP1 binding cell subpopulations are identified to be among ALDH+CD44+CXCR4+CD24+ cells. TAP1 significantly inhibited PCa growth both in vitro and in vivo. TAP1 significantly improved the anti-proliferation effect of the anti-androgens (Charcoal dextran-stripped serum (CDSS)+Bicalutamide, Enzalutamide) and chemotherapeutic agents (Abiraterone, Docetaxel, Etoposide) in vitro. TAP1 treatment shortens the length of telomeres in ALDH+CD44+CXCR4+CD24+ cells and significantly reduces the expression of Homeobox B9 (HOXB9) and TGF-β2. In conclusion, PSA−/lo PCa cell-specific targeting peptide (TAP1) that suppressed PCa cell growth both in vitro and in vivo and improved the drug sensitivities of anti-androgens and chemotherapeutic agents at least through shortening the length of telomere and reducing the expression of HOXB9 and TGF-β2. Therapeutic peptides that specifically target prostate cancer stem cell might be a very valuable and promising approach to overcome chemoresistance and prevent recurrence in patients with PCa. 相似文献
20.
SP62, a Viable Mutant of Bacteriophage T4D Defective in Regulation of Phage Enzyme Synthesis 总被引:22,自引:14,他引:8
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John S. Wiberg Steven Mendelsohn Virginia Warner Kathleen Hercules Cynthia Aldrich Judith L. Munro 《Journal of virology》1973,12(4):775-792
SP62 is a mutant of bacteriophage T4D that was discovered because it produces fewer phage than the wild type in the presence of 5-fluorodeoxyuridine. In the absence of phage DNA synthesis, SP62 solubilizes host DNA slower than normal; this may explain the sensitivity to 5-fluorodeoxyuridine. In Escherichia coli B at 37 C in the absence of drugs, SP62 makes DNA at a normal rate and the kinetics of appearance of phage are nearly normal. Under the same conditions, SP62 produces T4 lysozyme (gene e) at a normal rate until 20 min, but then produces it at twice the normal rate until at least 60 min. It has long been known that, when T4 DNA synthesis is blocked (DNA− state) in an otherwise normal infection, the synthesis of a number of early enzymes continues beyond the shutoff time of about 12 min seen in the DNA+ state, but still stops at about 20 min. We have termed the 12-min shutoff event S1 and the 20-min shutoff event S2. We show here that, in the DNA+ state, SP62 makes four early enzymes normally, i.e., S1 occurs. However, in the DNA− state (where S1 is missing), SP62 continues to make dCTPase (gene 56), dCMP hydroxymethylase (gene 42), and deoxynucleotide kinase (gene 1) for at least an hour; this results in production of up to 13 times the normal level of dCTPase at 60 min after infection, or 6 times the DNA− level. We conclude that SP62 is defective in the second shutoff mechanism, S2, for these three enzymes. In contrast, SP62 causes premature cessation of dTMP synthetase production in the DNA− state; the result is a twofold underproduction of dTMP synthetase. Autoradiograms of pulse-labeled proteins separated by slab-gel electrophoresis in the presence of sodium dodecyl sulfate show that a number of other T4 early proteins, including the products of genes 45, 46, and rIIA, are synthesized longer than normal by SP62 in the DNA− state. Few late proteins are made in the DNA− state, but in autoradiograms examining the DNA+ state there is little or no effect of the SP62 mutation on the synthesis of T4 late or early proteins. Circumstantial evidence is presented favoring a role for the gene of SP62 in translation of certain mRNAs. At very high temperatures (above 43 C) in the absence of drugs, phage production, but not DNA synthesis, is much reduced in SP62 infections relative to wild-type T4 infections; this temperature sensitivity is greater on E. coli CR63 than on E. coli B. This property has facilitated recognition of the SP62 genotype and aided in complementation testing and genetic mapping. A later publication will provide evidence that SP62 defines a new T4 gene named regA, which maps between genes 43 and 62. 相似文献