Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “BACTERIOPHAGE”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 55 records · Page 3Linked to original sources
PubMed2006

Isolation and characterization of the smallest bacteriophage P4 derivatives packaged into P4-size head in bacteriophage P2-P4 system.

Bacteriophage P4, a satellite phage of coliphage P2, is a very useful experimental tool for the study of viral capsid assembly and cos-cleavage. For an in vitro cos-cleavage reaction study of the P2-P4 system, new shortened and selectable markers containing P4 derivative plasmids were designed as a substrate molecules. They were constructed by swapping the non-essential segment of P4 DNA for either the kanamycin resistance (kmr) gene or the ampicillin resistance (apr) gene. The size of the genomes of the resulting markers were 82% (P4 ash8 delRI:: kmr) and 79% (P4 ash8 delRI:: apr) of the wild type P4 genome. To determine the lower limit of genome size that could be packaged into the small P4-size head, these shortened P4 plasmids were converted to phage particles with infection of the helper phage P2. The conversion of plasmid P4 derivatives to bacteriophage particles was verified by the heat stability test and the burst size determination experiment. CsCl buoyant equilibrium density gradient experiments confirmed not only the genome size of the viable phage form of shortened P4 derivatives, but also their packaging into the small P4-size head. P4 ash8 delRI:: apr turned out to be the smallest P4 genome that can be packaged into P4-sized head.

Bacteriophage P2↗
PubMed1976

Bacteriophage N3 of Haemophilus influenzae. II. Infection of transformable cells by bacteriophage DNA.

Transfection of H. influenzae with N3 bacteriophage DNA shows a dependence on concentration of DNA and a sigmoidal shape of uptake of DNA. The efficiency of transfection is decreased in the two types of recombination-defective strains of H. influenzae; rec1- and rec2-. The fact that N3 bacteriophage can code for its own recombination system but transfection is dependent on host recombation system allows the conclusion that the primary recombination during transfection with N3 phage DNA is mediated exclusively by the bacterial recombination system.

Bacteriophages↗
PubMed1952

The intracellular growth of bacteriophages. I. Liberation of intracellular bacteriophage T4 by premature lysis with another phage or with cyanide.

A method is described for liberating and estimating intracellular bacteriophage at any stage during the latent period by arresting phage growth and inducing premature lysis of the infected cells. This is brought about by placing the infected bacteria into the growth medium supplemented with 0.01 M cyanide and with a high titer T6 lysate. It was found in some of the later experiments that the T6 lysate is essential only during the first half of the latent period. Cyanide alone will induce lysis during the latter part of the latent period. Using this method on T4-infected bacteria it is found that during the first half of the latent period no phage particles, not even those originally infecting the bacteria, are recovered. This result is in agreement with the gradually emerging concept that a profound alteration of the infecting phage particle takes place before reproduction ensues. During the second half of the latent period mature phage is found to accumulate within the bacteria at a rate which is parallel to the approximately linear increase of intracellular DNA in this system. However, the phage production lags several minutes behind DNA production. When 5-methyltryptophan replaced cyanide as the metabolic inhibitor, similar results were obtained. The curves were, however, displaced several minutes to the left on the time axis. The results are compared with Latarjet's (16) data on x-radiation of infected bacteria and with Foster's data (18) concerning the effect of proflavine on infected bacteria. Essential agreement with both is apparent.

Bacteria↗
PubMed1979

Cloning of bacteriophage T5 DNA fragments in plasmid pBR322 and bacteriophage lambda gtWES.

Bacteriophage T5 was digested with the restriction endonucleases HindIII and EcoRI and the resulting fragments were inserted into the plasmid pBR322 and the bacteriophage lambda gtWES as vectors. Approx. 15% of the phage genome was recovered in recombinant clones. The recombinants were characterized by restriction analysis, DNA/DNA hybridization employing Southern blots, and ability to complement or recombine with amber mutants of T5. The results obtained allow revisions of the physical map of the T5 genome and partial correlation of the physical map with the genetic map.

Chromosome Mapping↗
PubMed1981

Partial exclusion of bacteriophage T2 by bacteriophage T4: an exclusion-resistant mutation in gene 56 of T2.

The early genes of bacteriophage T2 are partially excluded from the progeny of crosses between the related bacteriophages T2 and T4. This is due to complete exclusion from the progeny of six exclusion-sensitive sites in T2. A mutation [exr(56)1] in the sensitive site near T2 gene 56 renders the site partially resistant against exclusion. This paper describes the mapping of the exr(56)1 mutation. The mutation was mapped between two clusters of amber 56 mutations in T2, but mapping was not completely unequivocal. Additional evidence for location of exr(56)1 within gene 56 was provided by the decrease in the activity of the gene 56 product (dCTPase: EC 3.6.1.12) induced by T2 exr(56)1 strains. The location of exr(56)1 within an essential gene contradicts the exclusion model proposed by Russell & Huskey (1974).

Genes, Viral↗
PubMed1975

Escherichia coli deoxyribonucleic acid synthesis mutants: their effect upon bacteriophage P2 and satellite bacteriophage P4 deoxyribonucleic acid synthesis.

Escherichia coli C strains containing different deoxyribonucleic acid (DNA) synthesis mutations have been tested for their support of the DNA synthesis of bacteriophage P2 and its satellite phage P4. Bacteriophage P2 requires functional dnaB, dnaE, and dnaG E. coli gene products for DNA synthesis, whereas it does not require the products of the dnaA, dnaC, or dnaH genes. In contrast, the satellite virus P4 requires functional dnaE and dnaH gene products for DNA synthesis and does not need the products of the dnaA, dnaB, dnaC, and dnaG genes. Thus the P2 and P4 genomes are replicated differently, even though they are packaged in heads made of the same protein.

Coliphages↗
PubMed1978

Insertion of bacteriophage lambda into the deo operon of Escherichia coli K-12 and isolation of plaque-forming lambdadeo+ transducing bacteriophages.

A procedure has been devised to isolate plaque-forming lambda cI857S7 transducing bacteriophage which carry the internal promoter, P3, of the deo operon of Escherichia coli and the deoB and deoD genes, while lacking the deoP and cytP promoters of the same operon, in order to study, specifically, regulation at the P3 site. This has been accomplished by selecting for the insertion of bacteriophage lambda into the deoA gene in a strain deleted for the normal lambda attachment site (delta att lambda) and isolating from this lysogen lambda spi- and lambda EDTAr phage. Among these, lambda pdeoB+D+ phage were identified by their transducing abilities. From in vivo enzyme induction experiments performed on a delta deo strain lysogenized with such phage, they were shown to carry the P3 promoter while lacking the deoP and cytP promoters. A lambdapdeo B+D+ phage phage was used to lysogenize a deo+ delta att lambda strain, integration of lambda occurring within the region of homology, and, from a heat-induced lysate of this strain, a plaque-forming lambda+ phage carrying the complete deo operon was obtained. Phage lambda was also inserted into the deoB and deoD genes and into the tdk gene. By isolating lambdaspi- and lambdaEDTAr phage from the deo::(lambda) mutants and determining which bacterial genes they carried and whether they retained the int gene of lambda, it was found that lambda had inserted into deoD with the same orientation as lambda inserted into attlambda, whereas lambda inserted into deoA and deoB had the opposite orientation. Deletions extending from the site of lambda insertion into the bacterial chromosome were isolated by selecting for heat-resistant revertants. These confirmed the order of markers to be deo-serB-trpR-thr and also placed a locus, msp, determining sensitivity or resistance of male strains to male-specific phages, between trpR and thr. For some reason unknown, but which may be related to the orientation of the lambda prophages, short deletions rendering the bacterium Ser- Thr+ were of much lower frequency from the deoD::(lambda) lysogen than from the other two lysogens. From an examination of the residual deoD enzyme levels in deoB::(lambda) mutants, it was deduced that there may be two promoter sites within the deoB::(lambda) mutants, it was deduced that there may be two promoter sites within the deoB gene, transcription from one of these being sufficient to account for the noncoordinate nature of the induction of deoB and deoD gene products.

Chromosome Mapping↗
PubMed1981

Genetic transformation in Staphylococcus aureus: demonstration of a competence-conferring factor of bacteriophage origin in bacteriophage 80 alpha lysates.

A virion component that is responsible for conferring competence to Staphylococcus aureus was demonstrated in lysates of bacteriophage 80 alpha, a serological group B phage. Isolated particles of 80 alpha could not be shown to confer significant levels of competence. The phage component had a density of about 1.3 g/cm3, was inactivated by pronase, and was inhibited by antiserum prepared against isolated infectious particles of a serological group B phage. Centrifugation through a Ficoll gradient resulted in separation of competence-conferring activity and plaque-forming units. It is concluded that this proteinaceous subvirion component constitutes a bona fide competence factor of bacteriophage origin.

Bacterial Proteins↗
PubMed1966

Bacteriophage-induced functions in Escherichia coli K (lambda) infected with rII mutants of bacteriophage T4.

Rutberg, Blanka (Karolinska Institutet, Stockholm, Sweden), and Lars Rutberg. Bacteriophage-induced functions in Escherichia coli K(lambda) infected with rII mutants of bacteriophage T4. J. Bacteriol. 91:76-80. 1966.-When Escherichia coli K(lambda) was infected with rII mutants of phage T4, deoxycytidine triphosphatase, one of the phage-induced early enzymes, was produced at initially the same rate as in r(+)-infected cells. Deoxyribonuclease activity was one-third to one-half of that of r(+)-infected cells. This lower deoxyribonuclease activity was observed also in other hosts or when infection was made with rI or rIII mutants. Presence of chloramphenicol did not allow a continued synthesis of phage deoxyribonucleic acid in rII-infected K(lambda). No phage lysozyme was detected nor was any antiphage serum-blocking antigen found in rII-infected K(lambda). It is suggested that the rII gene is of significance for the expression of phage-induced late functions in the host K(lambda).

Antigens↗
PubMed1975

Escherichia coli capsule bacteriophages. III. Fragments of bacteriophage 29.

A glycanase activity, catalyzing the depolymerization of host capsular polysaccharide, is associated with Escherichia coli capsule bacteriophage no. 29, a small virus with an isometric head, carrying a base plate with a set of spikes. The bacteriophage particles were disrupted by mild acid treatment (5 to 8 min at pH 3.5 and 37 C), and the enzymatically active fragments were isolated and subjected to sodium dodecyl sulfate-gel electrophoresis as well as to electron microscopy. Of the at least nine different polypeptide chains found in the complete virion, three (of 57,000 plus or minus 3,000, 29,500 plus or minus 2,000 and 13,500 plus or minus 1,000 daltons) were detected in detached base plates. They had the appearance of six-pointed stars of about 14 nm in outer diameter, with a central hole or prop, carrying six (or, possibly, a multiple thereof) spikes. Two sizes of polypeptide chains (57,000 and 29,500) were found in pure spikes, cylindrical particles of about 14.5 to 15 nm in length and 5 nm in diameter, and one (57,000) in -- still capsule depolymerizing -- spike subunits of roughly 5 nm in diameter. Phage 29 spike preparations, homogeneous in analytical ultracentrifugation and immunoelectrophoresis, were found to have a molecular weight of 245,000, as determined from the sedimentation equilibrium, and to contain equimolar amounts of the two polypeptides, probably three copies of each per organelle. The amino acid analysis of the isolated spikes revealed that aspartic acid, alanine, serine, and glycine are their dominant constituents; no amino sugars or other carbohydrates were detected in the preparations.

Amino Acids↗
PubMed1975

Complete and Defective Bacteriophages of Classical Vibrio cholerae: Relationship to the Kappa Type Bacteriophage.

The Classical Vibrio cholerae strain NIH 41 contains two temperate bacteriophages, designated VcA-1 and VcA-2, that are distinguished by immunity, plaque morphology, induction kinetics, and particle morphology. Both phage are serologically related to phage Kappa. However, only phage VcA-2 has the Kappa type host range and immunity. The induction kinetics and immunity patterns of Classical vibrios suggest that these strains may contain defective phage related to the phages isolated from NIH 41. Classical strain 569B releases phage-tail structures upon induction that are morphologically and serologically related to both phages VcA-1 and VcA-2. The possible reason for the defectiveness of these phages in 569B is discussed. It is concluded that complete or defective bacteriophages of the Kappa type morphology and serology are extremely prevalent in V. cholerae, regardless of biotype.

Journal Article↗
PubMed1968

Replication of bacteriophage ribonucleic acid: analysis of the ultrastructure of the replicative form and the replicative intermediate of bacteriophage R17.

A detailed qualitative and quantitative comparison was made of the ultrastructure of single-stranded ribonucleic acid (RNA) from bacteriophage R17 and double-stranded replicative form (RF) and replicative intermediate (RI) from cells infected with this bacteriophage. The nucleic acids were prepared for electron microscopy by the protein monolayer spreading technique of Kleinschmidt. Single-stranded RNA aggregated during spreading in the absence of urea, whereas RF and RI did not. On the other hand, RF and RI appeared to be susceptible to shear during spreading, whereas R17 RNA was not. From the maximal length of RF, a base translation of 3.14 A was calculated. This value favors a 10-fold helix model of double-stranded RNA. The same base translation was found for R17 RNA, indicating a stacked base structure for single-stranded RNA spread in the presence of urea. RI is a branched structure and the branches are removed by ribonuclease treatment. The branches are believed to be nascent single-stranded viral RNA. The contour length of the branch was equal to the contour length of the main chain up to the branch point, as predicted from theoretical analysis of the replication of viral RNA. The structure of RF and the main chain of RI was also analyzed by plotting the log (end-to-end distance squared) versus log (contour length). This demonstrated structures intermediate in stiffness between a random coil and a rigid rod.

Coliphages↗
PubMed1979

Transduction of bacteriophage lambda by bacteriophage T1.

When bacteriophage T1 was grown on bacteriophage lambda-lysogenic cells, phenotypically mixed particles were formed which had the serum sensitivity, host range, and density of T1 but which gave rise to lambda phage. T1 packaged lambda genomes more efficiently both when the length of the prophage was less than that of wild-type lambda and when the host cell was polylysogenic. Expression of the red genes of lambda or the recE system of Escherichia coli during T1 growth enhanced pickup of lambda by T1, whereas packaging was reduced in recB cells. If donors were singly lysogenic, the expression of transduced lambda genomes as a PFU required lambda-specified excisive recombination, whereas lambda genomes transduced from polylysogens required only lambda- or E. coli-specified general recombination to give a productive infection.

Coliphages↗
PubMed1980

Control of early gene expression of bacteriophage T4: involvement of the host rho factor and the mot gene of the bacteriophage.

Many early mRNA species of bacteriophage T4 are not synthesized after infection of Escherichia coli in the presence of chloramphenicol. This has been interpreted as a need for T4 protein(s) to be synthesized to allow expression of some early genes, e.g., those for deoxycytidinetriphosphatase, deoxynucleosidemonophosphate kinase and UDP-glucose-DNA beta-glucosyltransferase. In the experiments described here, early mRNA of bacteriophage T4 was allowed to accumulate during chloramphenicol treatment. After the addition of rifampin to inhibit further RNA synthesis, and subsequent removal of chloramphenicol, the accumulated mRNA was permitted to express itself into measured enzyme activities. It was shown that the early mRNA species coding for deoxycytidinetriphosphatase and UDP-glucose-DNA beta-glucosyltransferase could be formed in the presence of chloramphenicol if the E. coli host cell carried a mutation in the structural gene for the RNA chain termination factor rho. This was interpreted to mean that T4 protein(s) with anti-rho activity is normally required for the expression of these two early genes. An altered rho-factor could not, however, relieve the need of phage protein synthesis for the formation of another early mRNA, that coding for deoxynucleosidemonophosphate kinase. In this case the mot gene of T4 seemed to be involved, since the primary infection of E. coli cells with the mot gene mutant tsG1 did not allow subsequent deoxynucleoside monophosphate kinase mRNA synthesis after wild-type phage infection in the presence of chloramphenicol. In control experiments, deoxynucleoside monophosphate kinase mRNA synthesis induced by wild-type phage superinfecting in the presence of chloramphenicol was facilitated by the primary infection with T4 phage containing an unmutated mot gene.

Chloramphenicol↗
PubMed1980

Derivation of a restriction map of bacteriophage T3 DNA and comparison with the map of bacteriophage T7 DNA.

The DNA of bacteriophage T3 was characterized by cleavage with seven restriction endonucleases. AvaI, XbaI, BglII, and HindIII each cut T3 DNA at 1 site, KpnI cleaved it at 2 sites, MboI cleaved it at 9 sites, and HpaI cleaved it at 17 sites. The sizes of the fragments produced by digestion with these enzymes were determined by using restriction fragments of T7 DNA as molecular weight standards. As a result of this analysis, the size of T3 DNA was estimated to be 38.74 kilobases. The fragments were ordered with respect to each other and to the genetic map to produce a restriction map of T3 DNA. The location and occurrence of the restriction sites in T3 DNA are compared with those in the DNA of the closely related bacteriophage T7.

Base Sequence↗
PubMed1981

Dual functions of bacteriophage T4D gene 28 product: structural component of the viral tail baseplate central plug and cleavage enzyme for folyl polyglutamates. II. Folate metabolism and polyglutamate cleavage activity of uninfected and infected Escherichia coli cells and bacteriophage.

We investigated the role of the T4D bacteriophage gene 28 product in folate metabolism in infected Escherichia coli cells by using antifolate drugs and a newly devised assay for folyl polyglutamate cleavage activity. Preincubation of host E. coli cells with various sulfa drugs inhibited phage production by decreasing the burst size when the phage particles produced an altered gene 28 product (i.e., after infection under permissive conditions with T4D 28(ts) or T4D am28). In addition, we found that another folate analog, pyrimethamine, also inhibited T4D 28(ts) production and T4D 28am production, but this analog did not inhibit wild-type T4D production. A temperature-resistant revertant of T4D 28(ts) was not sensitive to either sulfa drugs or pyrimethamine. We developed an assay to measure the enzymatic cleavage of folyl polyglutamates. The high-molecular-weight folyl polyglutamate substrate was isolated from E. coli B cells infected with T4D am28 in the presence of labeled glutamic acid and was characterized as a folate compound containing 12 to 14 labeled glutamate residues. Extracts of uninfected bacteria liberated glutamate residues from this substrate with a pH optimum of 8.4 to 8.5. Extracts of bacteriophage T4D-infected E. coli B cells exhibited an additional new folyl polyglutamate cleavage activity with a pH optimum of about 6.4 to 6.5, which was clearly distinguished from the preexisting activity in the uninfected host cells. This new activity was induced in E. coli B cells by infection with wild-type T4D and T4D amber mutants 29(-), 26(-), 27(-), 51(-), and 10(-), but it was not induced under nonpermissive conditions by T4D am28 or by T4D 28(ts). Mutations in gene 28 affected the properties of the induced cleavage enzyme. Wild-type T4D-induced cleavage activity was not inhibited by pyrimethamine, whereas the T4D 28(ts) activity induced at a permissive temperature was inhibited by this folate analog. Folyl polyglutamate cleavage activity characteristic of the activity induced in host cells by wild-type T4D or by T4D gene 28 mutants was also found in highly purified preparations of these phage ghost particles. The T4D-induced cleavage activity could be inhibited by antiserum prepared against highly purified phage baseplates. We concluded that T4D infection induced the formation of a new folyl polyglutamate cleavage enzyme and that this enzyme was coded for by T4D gene 28. Furthermore, since this gene product was a baseplate tail plug component which had both its antigenic sites and its catalytic sites exposed on the phage particle, it was apparent that this enzyme formed part of the distal surface of the phage baseplate central tail plug.

Carboxypeptidases↗
PubMed1984

Bacteriophage T4 bypass31 mutations that make gene 31 nonessential for bacteriophage T4 replication: isolation and characterization.

T4 bacteriophage mutants called bypass31 (byp31) that specifically suppress gene 31 amber mutations have been isolated and characterized. The mechanism by which the byp31 mutation, byp31-1, suppresses gene 31 nonsense mutations does not involve synthesis of gp31 or of a particular gp31 fragment; furthermore, the byp31 allele suppresses all nonsense mutations in gene 31 that have been tested. We detect no unusual properties among the T4 particles made in su- cells by the T4amN54byp31-1 double mutant. These virions, made in the absence of gp31, show normal heat sensitivity, normal sensitivity to osmotic shock, and normal morphology. Specific different gene 31 missense mutants are able to form plaques with high efficiencies on the following two types of host defective cells: (i) Escherichia coli groEL (Tilly et al., Proc. Natl. Acad. Sci. U.S.A. 78:1629-1633, 1981) mutants that block T4 capsid assembly and (ii) E. coli rho mutants in which T4+ heads are assembled, but in which tail production and DNA synthesis are blocked. (Note that not all rho mutants block T4 production [G. Binkowski and L. D. Simon, p. 342-350, in C. K. Mathews, E. M. Kutter, G. Mosig, and P. B. Berget, ed., Bacteriophage T4, 1983]; T4 is able to replicate in rho mutants such as rho ts15, whose principal defect is that they fail to terminate transcription.) The byp31-1 allele permits production of T4 particles in E. coli groEL host-defective mutants, but not in E. coli rho host mutants.

DNA Replication↗