equimolar addition of oligoribonucleotides with t4 rna ligase.t4 induced rna ligase will join equimolar concentrations of two oligoribonucleotides, (ap)3c and p(up) 5, to form a single product, (ap)3cp(up) 5, in high yield. the presence of the 3' phosphate on p(up)5 prevents the oligomer from adding to itself. the ph optimum of the reaction is about 7.5, but less of the undesirable adenylated intermediate, app(up) 5, forms at ph 8.2. the reaction rate is a linear function of oligomer concentration from 3 micronm to 0.6 mm. the data suggest that t4 rna lig ...197717097
replication of the linear mitochondrial dna of tetrahymena pyriformis.1. electron micrographs of the linear mtdna from tetrahymena pyriformis strain gl show linear molecules with a duplex 'eye' of variable size in the middle. this indicates that replication of this dna starts near the middle of the molecule and proceeds bidirectionally to the ends, as previously shown for the mtdna of strain st (arnberg, a.c., van bruggen, e.f.j., clegg, r.a., upholt, w.b. and borst, p. (1974) biochim. biophys. acta 361, 266-276). the mtdnas of these two strains have little base s ...1979110348
effect of chloramphenicol and starvation for an essential amino acid on the synthesis and decay of t4 bacteriophage-specific messengers transcribed from early and quasi-late promoters. 1975170861
phosphorylation of double-stranded dnas by t4 polynucleotide kinase.the phosphorylation by t4 polynucleotide kinase of various double-stranded dnas containing defined 5'-hydroxyl end group structures has been studied. particular emphasis was placed on finding conditions that allow complete phosphorylation. the dnas employed were homodeoxyoligonucleotides annealed on the corresponding homopolymers, dna duplexes corresponding to parts of the genes for alanine yeast trna, and a suppressor tyrosine trna from escherichia coli. the rate of phosphoylation of dnas with ...1976178357
slow switchover from host rna synthesis to bacteriophage rna synthesis after infection of escherichia coli with a t4 mutant defective in the bacteriophage t4-induced unfolding of the host nucleoid.most, if not all, host rna synthesis was shut off after infection of escherichia coli strain b/5 with a bacteriophage t4 multiple mutant defective in the abilities to induce (i) unfolding of the host nucleoid (unf-), (ii) nuclear disruption (ndd-), and (iii) host dna degradation (dena-, denb-). the shutoff of host rna synthesis and turn-on of phage rna synthesis were slower after infection of e. coli with unf- phage than after infection with unf+ phage. this delay in the switchover from host rna ...1977201776
regulation of ribosome function following bacteriophage t4 infection. 1978210283
the purification of nuclease-free t4-rna ligase.rna ligase has been highly purified in good yields from bacteriophage t4-infected escherichia coli by a rapid and reproducible procedure. the enzyme is free of phosphomonoesterase and ribonuclease activities and is therefore suitable for the synthesis of oligoribonucleotides and for the labeling of the 3'-terminus of rna. greater than 90% of the protein in the enzyme preparation migrates as a single band on gradient polyacrylamide gels containing sodium dodecyl sulfate during electrophoresis. fo ...1979219895
a new system for studying molecular mechanisms of mutation by carcinogens.a new system for studying the molecular mechanisms of mutation by carcinogens is described. the system involves (a) site-specific modification of the essential gene g in phi x174 replicative form dna by a combination of chemical and enzymatic steps; (b) production of mutant virus carrying a change at a single preselected site by transfection of spheroplasts with the site modified phi x174 dna; (c) detection and propagation of mutants using a host carrying the plasmid, p phi xg, that rescues all ...1979227905
mutants of escherichia coli "cryptic" for certain periplasmic enzymes: evidence for an alteration of the outer membrane.mutants in which the expression of periplasmic enzymes by whole cells is reduced (termed "cryptic") are also found to show greatly reduced uptake of labeled adenosine 5'-monophosphate (5'-amp), providing a rapid assay for crypticity. the crypticity of 3'- and 5'-nucleotidase has been examined as a function of substrate concentration. the km for 3'- or 5'-amp increases in the cryptic mutants when whole cells are used as the enzyme source. the vmax is not altered. electrophoretic analysis of prote ...1977320175
isolation and characterization of conditional-lethal rho mutants of escherichia coli.temperature-sensitive nita (rho) mutants of e. coli were isolated; one of them was characterized as an amber mutant. these strains show the nit phenotype (transcription of phage lambda dna independent of the n gene) at low temperatures and are inviable at high temperatures. the mutated sites appear to be between cya and mete on the chromosome. temperature-sensitive nita bacteria not only permit leftward transcription of the lambda genome at a high rate in the absence of the lambda n protein, but ...1977322147
regulation of the expression of bacteriophage t4 genes 32 and 43. 1977324117
proteolysis during t4 head maturation: evidence against the involvement of a phage-induced trypsin-like enzyme. 1977327685
t4 phage-coded deoxycytidylate hydroxymethylase: purification and studies on intermolecular interactions. 1977332172
stimulation of cell division by t4 infection in escherichia coli dnaets at nonpermissive temperature.filamentous cells resulting from growth of a dnaets mutant of escherichia coli at high temperature were stimulated to divide by infection with bacteriophage t4. the effect appears to be related to t4 dna synthesis; no increase in cell number took place in chloramphenicol-treated. t4-infected cells nor in cells infected with dna synthesis-less mutants of t4. the ability of cells to divide after t4 infection was dependent on the length of time that the cells had been grown at 42 degrees c, indicat ...1977337113
attachment of tail fibers in bacteriophage t4 assembly. purification, properties, and site of action of the accessory protein coded by gene 63. 1978344316
construction and properties of recombinant plasmids containing the rii genes of bacteriophage t4.the ecori digestion products of phage t4 dna have been examined using a phage dna transformation assay. a 2.6 x 10(6) dalton fragment was found to contain the rii genes. this fragment was purified and then treated with hindiii endonuclease. the cleavage products were ligated to the vector plasmid pbr313 and viable recombinant plasmids recovered. a genetic assay was employed to demonstrate that the recombinants contained t4 dna and to localize on the phage genetic map the ecori and hindiii sites ...1978345100
genetic and physiological characterization of escherichia coli k12 mutants (tabc) which induce the abortive infection of bacteriophage t4. 1978351929
selectivity of rna chain initiation in vitro. 1. analysis of rna initiations by two-dimensional thin-layer chromatography of 5'-triphosphate-labeled oligonucleotides.a method for the rapid and quantitative analysis of 5'-terminal oligonucleotides of rnas made in vitro is described. the method involves synthesis of rna in the presence of [gamma-32p]atp or gtp, isolation of the rna, and digestion with t1 or pancreatic ribonucleases to release labeled 5'-triphosphate termanated oligonucleotides. the oligonucleotides are then subjected to chromatography on a polyethyleniminecellulose thin-layer system using 2 m licl, 0.01 m edta (ph 6.5) in the first dimension a ...1978352390
ionic strength perturbation kinetics of gene 32 protein dissociation from its complex with single-stranded dna.equilibrium and kinetic studies of the interaction of gene 32 protein of t4 phage with single-stranded fd dna were performed monitoring the changes in protein fluorescence. from the fluorescence titrations, it was estimated that a monomer of gene 32 protein covered six nucleotide bases on the dna and the lower limit for the apparent association constant was 1.9 x 10(8) m-1 with a cooperative parameter of 10(3) in 0.1 m 2-amino-2-hydroxymethyl-1,3-propanediol hydrochloride (ph 7) at 25 degrees c. ...1978359047
glutathione-dependent enzyme reactions of the phage t4 ribonucleotide reductase system. 1978359548
role of lipopolysaccharide and outer membrane protein of escherichia coli k-12 in the receptor activity for bacteriophage t4.lipopolysaccharide isolated from escherichia coli k-12 did not inactivate phage t4, although the cell envelopes with 1% sodium deoxycholate resulted in the release of cytoplasmic membrane proteins, 70% of the lipopolysaccharide, and almost all of the phospholipid. the reconstitution of phage receptor activity was achieved from deoxycholate-soluble and -insoluble fractions by dialysis against a solution of magnesium chloride. lipopolysaccharide was the only essential component in the deoxycholate ...1978361717
model for dna packaging into bacteriophage t4 heads.the mechanism of dna packaging into bacteriophage t4 heads in vivo was investigated by glucosylation of hydroxymethylcytosine residues in a conditionally glucose-deficient host. cytoplasmic dna associated with partially packaged ts49 heads can be fully glucosylated, whereas dna already packaged into these heads is shown to be resistant to glucosylation. after temperature shift and completion of arrested packaging into the reversible temperature-sensitive ts49 head, the structure of the dna in th ...1978364076
[selective binding of oligoribonucleotides by t7 phage induced rna-polymerase].it was shown previously that e. coli rna-polymerase being incubated with the random oligonucleotide mixtures of definite length binds certain oligoribonucleotides with the length greater than or equal to 5 nucleotides. the data presented demonstrate that t7 phage induced rna-polymerase (t7 rna-polymerase) also binds selectively oligoribonucleotides beginning from pentaribonucleotides. from the random mixtures of penta-, hexa-, hepta-, octa-, nona- and decaribonucleotides the hepta- and octaribon ...1978370554
dna replication with bacteriophage t4 proteins. purification of the proteins encoded by t4 genes 41, 45, 44, and 62 using a complementation assay.the proteins encoded by bacteriophage t4 genes 41, 45, 44, and 62 are known from the genetic studies of epstein et al. ((1963) cold spring harbor symp. quant. biol. 28, 375--394) to be required for viral dna synthesis. a convenient assay for each of these proteins is described which is based on the specific stimulation by each protein of dna synthesis in extracts of escherichia coli infected with mutants of bacteriophage t4 unable to make that protein. the t4 41 protein, 45 protein, and the comp ...1979376524
restriction in vivo. iii. general effects of glucosylation and restriction on phage t4 gene expression and replication. 1979380145
[asymmetry in the frequencies of reciprocal recombinants in crosses of t4 phage riib mutants].the frequencies of reciprocal recombinants in crosses between riib mutants of t4 phage were shown to differ from each other. in terms of the correction model, this asymmetry of genetic recombination was used to measure the comparative correctability of the mismatched regions to the wild type and to the mutant alleles. the data obtained are in quantitative agreement with the analogous values for the same mismatched regions determined by comparison of the markers located at the same site. this str ...1979391644
nucleotide sequence of the region required for maintenance of colicin e1 plasmid.plasmids carrying various portions of colicin e1 plasmid (cole1) dna have been isolated in an attempt to determine the regions of cole1 dna which are required for maintenance of the plasmid in bacteria. to construct the plasmids, the dna of a cole1 derivative that contains a gene which controls ampicillin resistance was cleaved by the restriction endonuclease haeii. the digestion products were joined by t4 dna ligase and then used to transform bacteria to ampicillin resistance. the plasmid deriv ...1979393952
platinum(ii) complexes block the entry of t4 phage dna into the host cells.the efficiency of multiplicity reactivation of t4 particles inactivated by platinum(ii) complexes is very low. the same is true for marker rescue and functional survival of genes. this can be at least partly explained by the inability of most inactivated virus particles to introduce their dna into the host cells as demonstrated by electron microscopy. conformational changes in the dna, formation of dna-dna and dna-protein cross-links and the damage of proteins participating in the injection proc ...1979398750
[interaction between alkaloids of claviceps purpurea and development of some bacteria and bacteriophages. preliminary experiments].it has been tested the bacteriolytic activity and the interaction among the development of the bacteriophage t4 and ergot alkaloids. it has been determined a bacteriolytic action on the bacterial stub "e. coli host of bacteriophage t4. it exist an interaction also among such alkaloids and the development of the bacteriophage t4, which appears through an increase of the quantity of lysis areas, becoming evident in solid bodies containing the lysing bacterium. further researches with other bacteri ...1979400107
genetic control of bacteriophage t4 baseplate morphogenesis. i. sequential assembly of the major precursor, in vivo and in vitro. 1975765481
[effect of restricting the action of an amber-mutation suppressor contained in bacteriophage t4 genome].the action of a bacteriophage suppressor can be restricted due to mutations arising in the genome of the host bacteria. bacterial strains escherichia coli bn and can were isolated in which a complete restriction of the action of phage suppressor psu+ took place. in thees strains obtained the restriction of serin-specific phage suppressors psu + a and psu + b is brought about. the action of bacteriophage suppressor su3+ containing in e. coli can is not abolished in this strain. the abolish of the ...1975767202
studies on phage internal proteins: formation of internal protein - t2 dna complexes in vivo.internal proteins, synthesized in t2-infected escherichia coli b cells were recovered from bacterial membranes during the early stages of infection. approx. 15 min after the onset of infection, t2 and t4 internal proteins were released from the bacterial membranes and sedimented along with newly synthesized phage dna. internal protein-dna complexes were also obtained by chromatography on hydroxylapatite columns. internal proteins were not released from bacterial membranes after infection with am ...1976772169
a study of possible mechanisms of the rna-polymerase involement in mutagenesis in phage t4.spontaneous and induced mutation frequencies of phage t4 have been measured in escherichia coli strains containing altered rna-polymerase. in the strain e. coli rif-r stl-r, with double rna-polymerase mutation, spontaneous reversion rates were increased in different mutants of phage t4. the study of base analogues mutagenesis in ruv mutants of phage t4 has shown that the introduction of rna-polymerase mutations did not increase reversion rates in a mutant of frame-shift type but enhanced the rat ...1976775319
induced mutagenesis in bacteriophage t4 growing in strains of e. coli with altered rna-polymerase.enhanced reversion frequencies of t4 r mutants in e. coli strains with altered rna polymerase have been obtained. the results reported have confirmed previous data on the effect of rna-polymerase on the process of mutagenesis [2]. no such effect has been found in the course of studies of the recombination process.1976775324
lipopolysaccharide-deficient, bacteriophage-resistant mutants of escherichia coli k-12.bacteriophage-resistant mutants isolated and classified in a previous study were examined for alterations in their lipopolysaccharide (lps) composition, and properties likely to be affected by alterations in lps composition were studied. it was found that many of the mutants of the ktw (k2-resistance), ttk (t2, t4, or k19 resistance), bar (bacteriophage), wrm (wide-range mutants), and miscellaneous resistance groups were altered in their response to a series of antibiotics and to two lps-specifi ...1976776951
purification of a proteolytic enzyme from t4-infected escherichia coli cells. 1976779237
t4 dna injection. ii. protection of entering dna from host exonuclease v. 1976779243
properties and structure of a gene 24-controlled t4 giant phage. 1976781276
replication of bacteriophage t4 dna in vitro. i. basic properties of the system.a new in vitro system for t4 dna replication was developed by concentrating cell lysates on cellophane disks. the time course of [3h]dttp incorporation into dna by the system was separated into two phases: one was a very rapid incorporation which was terminated within 2 min (phase i reaction), and the other was a slow but continuous incorporation thereafter (phase ii reaction). more than half of the phase i reaction product was escherichia coli dna, but the phase ii reaction was mostly t4 dna. p ...1976785023
[biology of spheroplast- and protoplast-like types of l-forms of escherichia coli k12 converted with penicillin].a total of 21 strains of stable l-forms were obtained under the action of penicillin on various hfr and f- strains of e. coli k12. three morphological types of the l-forms obtained differed by the character of the cell elements, sensitivity to chemical agents, antibiotics and to t4 and t6 phages. cell wall was revealed in one type of the l-forms, but the rest l-form types were devoid of the cell walls. reference of the l-forms which preserved the cell wall to the spheroplastic type, and the l-fo ...1976795246
cleavage of t4-induced proteins during phage morphogenesis: characterization of peptides.polypeptides of low mol. wt. have been extracted from t4 coliphages and from escherichia coli b cells infected with a wild type and various amber mutants of bacteriophage t4. six peptides were fractionated by chromatography on phosphocellulose: three of them were cleaved from proteins synthesized late in infection and related to phage head. the remaining three peptides have been shown to arise from early-labelled phage-induced proteins. two of these six small petide fragments were found in the h ...1976778336
dna crosslinks, single-strand breaks and effects on bacteriophage t4 survival from tritium decay of (2-3h)adenine, (8-3h)adenine and (8-3h)guanine. 1976772217
mechanism localisation and control of restriction cleavage of phage t4 and lambda chromosomes in vivo.the primary action of restriction endonuclease, cleaving infecting dna, has been demonstrated in vivo. this primary cleavage is followed rapidly by hydrolysis of the cleaved dna at its newly exposed termini. infecting viruses can inactivate cytoplasmic and membrane restriction endonucleases to prevent cleavage of unmodified dna replicas.1976768782
purification and some properties of deoxyribonuclease whose synthesis is controlled by gene 49 of bacteriophage enzyme which specifically cleaves very-fast-sedimenting dna of bacteriophage t4 is synthesized after infection of t4, and its synthesis is controlled by gene 49 [1,2]. this enzyme has been proved to be a dnase [2]. we have purified this dnase 3000-fold from extracts of e. coli infected with t4. the purified preparation was practically free from other dnases, and the dnase activity was not detectable in cells infected with a mutant defective in gene 49. the enzyme activity from cells infected ...1979389625
human globin messenger rna: importance of cloning for structural analysis.the sequence of most of the human beta globin messenger rna and large sections of the alpha globin messenger rna has been determined. partly because of genetic polymorphism, it was necessary to clone globin complementary dna in order to extend the analysis. purified human fetal globin messenger rna was isolated and used as a template by reverse transcriptase to produce duplex complementary dna molecules. these molecules were linked in vitro to plasmid dna by use of t4 ligase in the presence of e ...1977847468
the effect of template secondary structure on vaccinia dna polymerase.vaccinia virus dna polymerase will utilize a substrate consisting of phi x174 dna primed with a strand of a unique restriction fragment, but the reaction is inefficient. examination of the reaction products by alkaline agarose gel electrophoresis revealed a few discrete fragments, each corresponding to an extended primer strand. this result implies that specific barriers exist on the phi x174 template which impede, but do not completely halt, the progress of the enzyme. only a few per cent of th ...1979381293
[topological model of bacteriophage-t4 dna replication].the structure and function of the dna--membrane complex in e. coli cells infected with bacteriophage t4 was studied. the dna--membrane complex was isolated from the cells pulse or uniformly labeled with 3h-thymidine, fractionated by detergent treatment and separated on a discontinuous sucrose gradient. the attachment of small dna fragments to the plasma membrane was analysed. replicating bacteriophage t4 dna was reversibly associated with the cytoplasmic membrane in the wall/membrane adhesion zo ...1977377048
s1 nuclease as a probe for the conformation of a dimeric trna precursor.we have employed s1 nuclease to probe the structure of an intermediate in trna biosynthesis available only in radiochemical purity. the dimeric precursor to trnagln and trnaleu from bacteriophage t4 was digested with the single-strand specific nuclease, and the products of the reaction were compared with the s1 digestion products of the mature cognate trna's. quantitation and sequence analysis of the products revealed that the location and accessibility of s1 cleavage sites in the precursor were ...1979369598
construction and properties of a cell-free system for bacteriophage t4 late rna synthesis.a cell-free system for synthesizing bacteriophage t4 late rna is described. the system, which is based on the "cellophane disc" technique introduced by schaller and co-workers (schaller, h., otto, b., nüsslein, v., huf, j., hermann, r., and bonnhoeffer, f. (1972) j. mol. biol. 63, 183-200), provides favorable conditions for t4 dna and rna synthesis in vitro. total rna synthesis can be sustained for more than 1 h at 25 degrees c and initiation of early and late rna chains occurs in vitro. the cap ...1979368054
comparison of the effects of bacteriophage t4 infection and n-ethylmaleimide on the translational specificity of escherichia coli ribosomes. 19751091211
transcription of bacteriophage t4 genome in vitro. heterogeneity of rna polymerase in crude extracts of normal and t4-infected escherichia coli order to obtain rna polymerase preparations carrying the necessary specificity determinants to transcribe the delayed-early genes of bacteriophage t4, crude extracts of uninfected and t4-infected escherichia coli were fractionated in glycerol gradients of low ionic strength. in contrast to the reported sedimentation behavior of the purified enzyme, the rna polymerase activity in crude extracts of normal and infected cells sedimented heterogeneously over a wide range of sedimentation coefficie ...19751091288
an escherichia coli ribonuclease which removes an extra nucleotide from a biosynthetic intermediate of bacteriophage t4 proline transfer rna.the biosynthesis of bacteriophage t4 trnapro, trnaser, and trnaile requires enzymatic removal of extra nucleotides from the 3' terminus of the respective precursor rnas. a ribonuclease activity capable of catalyzing such reactions has been partially purified from uninfected escherichia coli using an artificial precursor rna as substrate. a number of ribonuclease activities were resolved during purification. use of e. coli strain bn, a mutant known to be deficient in the relevant ribonuclease act ...1978364422
further characterization of the r plasmid rts1 and its mutant ptw2: replication and incompatibility of the plasmid.incompatibility of the r plasmid rts1 and its replication mutant ptw2 was studied in reca host cells of escherichia coli. when the r plasmid r401, belonging to the same incompatibility group as rts1, was used as a test plasmid, r401 was eliminated preferentially from (rts-r401)+ cells irrespective of the direction of transfer. in contrast, ptw2 and r401 were mutually excluded. the decreased incompatibility of ptw2 was confirmed by a direct incompatibility test in which a derivative of rts1 expel ...1978355218
proceedings: modification of e. coli rna polymerase induced by t4 phage infection. 19751094019
functional compartmentation of dna precursors in t4 phage-infected bacteria. 1978348692
packaging of dna into t4 bacteriophage: exclusion of host dna despite the absence of both host dna degradation and nuclear disruption. 19751096456
recovery of polysome function of t4-infected escherichia coli after brief treatment with chloramphenicol and rifampin.t4-infected escherichia coli cells briefly exposed to rifampin, or to rifampin plus chloramphenicol, were capable of protein synthesis for some time after removal of the antibiotics, although ribonucleic acid synthesis was irreversibly inhibited. partially completed peptides trapped on polysomes by high levels of chloramphenicol were eventually completed after removal of the drug, as demonstrated by subjecting labeled peptides from appropriate polysome regions to polyacrylamide disc gel electrop ...19751096805
the nucleotide sequence of the dimeric precursor to glutamine and leucine transfer rnas coded by bacteriophage t4. 19751097716
carbon loss during irradiation of t4 bacteriophages and e. coli bacteria in electron microscopes. 19751097727
repetitive dna replication of the incomplete genomes of phage t4 petite particles.the genomes of petite t4 phage particles presumably cannot circularize because they are deficient for a significant terminal segment and hence not terminally redundant like normal t4 genomes. combined density- and 32p-labeling shows that the majority of such deficient dna molecules can nevertheless replicate their entire length. furthermore, the density-shift technique shows that replicated parental strands can exchange their partners for new light strands, indicating that noncircularized t4 dna ...19751099579
analysis of bacteriophage t4 chloramphenicol rna by dna-rna hybridization and by cell-free protein synthesis, and the effect of escherichia coli polarity-suppressing alleles on its synthesis. 19751100847
relaxation complexes of poasmid dna and protein. iii. association of protein with the 5' terminus of the broken dna strand in the relaxed complex of plasmid cole1.the location of the protein in the open circular dna form of the cole1 dna-protein relaxation complex, induced by treatment with sodium dodecyl sulfate, has been studied using several enzymes of dna metabolism. escherichia coli exonucleases i and iii are able to degrade extensively the nicked strand of the relaxed complex from the 3' end. dna polymerase i can initiate synthesis using the relaxed complex as template-primer and specifically extends the 3' end of the nicked strand. the 5' end of th ...19751102545
bacteriophage t4 virion dihydrofolate reductase: approaches to quantitation and assessment of function.this paper is concerned with the physiological role(s) of t4 phage-coded dihydrofolate reductase, which functions both in dna precursor metabolism and as a virion protein. (i) we have detected enzyme activity in noninfectious particles produced under restrictive conditions by gene 11 mutants. this supports the conclusion of kozloff et al. (j. virol. 16:1401-1408, 1975) that the protein lies in the baseplate, covered by the gene 11 protein. (ii) we have obtained further evidence for virion dihydr ...1977330880
effect of t4 modification of host valyl-trna synthetase on enzyme action in vivo. 19751103443
the repair of ultraviolet damage by phage t4: the role of the early phage genes. 19751103821
the biology of bacteriophage t4 transfer rnas. 19751104087
further studies on bacteriophage t4 dna synthesis in sucrose-plasmolyzed cells.this paper describes several technical improvements in the sucrose-plasmolyzed cell system used in earlier experiments on dna synthesis in situ with escherichia coli infected by dna-defective mutants of bacteriophage t4 (w. l. collinsworth and c. k. mathews, j. virol. 13:908-915, 1974). using this system, which is based primarily on that of m. g. wovcha et al. (proc. natl. acad. sci. u.s.a. 70:2196-2200, 1973), we reinvestigated the properties of mutants bearing lesions in genes 1, 41, and 62, a ...1977328926
phage t4-modified rna polymerase transcribes t4 late genes in vitro.initiation of t4 late rna synthesis has been achieved in an in vitro system prepared from escherichia coli cells infected with wild-type or maturation-defective mutant t4 phage. the system uses a cellophane membrane as a mechanical support for concentrated cell lysates and for added streptolydigin-resistant rna polymerases. transcriptional activity and selectivity of added rna polymerases are tested while endogenous rna polymerase activity is inhibited by streptolydigin. t4-modified rna polymera ...1977271954
excision of bromodeoxyuridine from t4-dna by an antimutator polymerase of t4 phage.with gene-43 (dna polymerase)-ts-mutants of t4 phage, l98 (mutator) and cb121 (antimutator), and the t4 wild type, double labelling of dna was carried out with (h3)-bromodeoxyuridine (budr) and (c14)-thymidine (tdr). experiments on (c14) tdr for dna synthesis measurement in the presence of budr offered evidence of the ability of the cb121 mutant to excise budr from the dna. this effect took place only at increased temperature. as distinct from dna synthesis of the host, all t4 phages used prefer ...1975239571
restoration by t4 ligase of dna sequences sensitive to "flush" cleaving restriction enzyme.fouteen "flush"-ended segments originate from the action of the restriction endonuclease hae iii of haemophilus aegiptius on the dna of the colicinogenic factor cole 1 (a. oka and m. takanami, nature, 264, 191, 1976). they are joined by the t4 polynucleotide ligase. the reaction can be monitored by gel electrophoresis, electron microscopy and resistance to phosphatase of the 5'-32p labelled ends. the joined products are a random recombination of the original segments, and can be cleaved by the s ...1977198743
site-specific mutagenesis using synthetic oligodeoxyribonucleotide primers: ii. in vitro selection of mutant dna.a method for the in vitro selection of mutant dna has been devised as an adjunct to the recently developed method for the use of short enzymatically-synthesized oligodeoxyribonucleotides of defined sequence as site-specific mutagens for circular dna. the selection method uses the mutating oligodeoxyribonucleotide as a primer for escherichia coli dna polymerase i (large fragment) under conditions where there is preferential interaction with mutant dna template. after ligation using t4 dna ligase, ...1979161246
role of polymeric forms of the bacteriophage phi x174 coded gene a protein in phi xrfi dna cleavage.gene a of the phi x174 genome codes for two proteins, a and a* (linney, e.a., and hayashi, m.n. (1973) nature new biol. 245, 6-8) of molecular weights 60,000 and 35,000, respectively. the phi x a* protein is formed from a natural internal initiator site within the a gene cistron while the phi x a protein is the product of the entire a gene. these two proteins have been purified to homogeneity as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis. previous studies have shown that ...1979158588
different specific activities of the monomeric and oligomeric forms of plasmid dna in transformation of b. subtilis and e. coli.(1) the low residual transforming activity in preparations of monomeric, supercoiled, circular (ccc) forms of the plasmids pc194 and phv14 could be attributed to the presence in such isolates of a small number of contaminating multimeric molecules. (2) e. coli derived preparations of phv14, as in vitro recombinant plasmid capable of replication in both e. coli and b. subtilis, contain oligomeric forms of plasmid dna in addition to the prevalent monomeric ccc form. the specific transforming activ ...1979113646
ligation-independent cloning of glutathione s-transferase fusion genes for expression in escherichia coli.a plasmid vector has been constructed that allows the ligation-independent cloning of cdnas in any reading frame and directs their synthesis in escherichia coli as glutathione s-transferase-linked fusion proteins. the cloning procedure does not require restriction enzyme digestion of the target sequence and does not introduce any additional sequences between the thrombin cleavage site and the foreign protein. extended single-stranded tails complementary between the vector and insert, generated b ...19921339364
revertants of double opal-mutants of bacteriophage t4.revertants of double opal-mutants of bacteriophage t4 have been obtained. the properties of these revertants suggest that reversion of double opal-mutants is effected by the activity of some gent-suppressor appeared in the phage genome. restriction of these revertants by streptomycin-resistant bacterial strains shows that the suppression of the opal-mutants is realized at translation.1976799254
the distribution of uv damage in the laci gene of escherichia coli: correlation with mutation spectrum.we have determined the uv (254 nm) damage distribution in the transcribed and non-transcribed strands of the i-d region of the escherichia coli laci gene. the locations of replication blocking lesions were revealed as termination sites of t7 dna polymerase and/or t4 dna polymerase 3'-5' exonuclease. termination products, i.e. both cyclobutane pyrimidine dimers and 6-4 photoproducts, were resolved and analysed on an automated dna sequencer. these two major photoproducts are not randomly distribut ...19921383713
roles of cys148 and asp179 in catalysis by deoxycytidylate hydroxymethylase from bacteriophage t4 examined by site-directed mutagenesis.the proposed roles of cys148 and asp179 in deoxycytidylate (dcmp) hydroxymethylase (ch) have been tested using site-directed mutagenesis. ch catalyzes the formation of 5-(hydroxymethyl)-dcmp, essential for dna synthesis in phage t4, from dcmp and methylenetetrahydrofolate. ch resembles thymidylate synthase (ts), an enzyme of known three-dimensional structure, in both amino acid sequence and the reaction catalyzed. conversion of cys148 to asp, gly, or ser decreases ch activity at least 10(5)-fold ...19921420151
the sucrose gradient and native dna s20,w, an examination of measurement problems.sedimentation coefficients of t7, t2h and t4 dna were determined with isokinetic sucrose gradients in both 0.1 m and 1 m nacl. the s values were completely equivalent to those measured by analytical ultracentrifuge and no reduction of s20,w was observed due to the presence of sucrose (anomalous sedimentation). s20,w values are calculated on the basis of both partial specific volume (v) and apparent specific volume (0). using the latter value s20,w molecular weight relations are derived for 0.1 m ...1976793631
molecular characterisation of a dna ligase gene of the extremely thermophilic archaeon desulfurolobus ambivalens shows close phylogenetic relationship to eukaryotic ligases.a 3382 bp fragment containing a gene for a dna ligase from the extremely thermophilic, acidophilic, and facultatively anaerobic archaeon (archaebacterium) desulfurolobus ambivalens was cloned and sequenced. the deduced amino acid sequence (600 amino acids, 67619 molecular weight) showed 30-34% sequence identity with the atp-dependent eucaryal (eukaryotic) dna ligases of schizosaccharomyces pombe, saccharomyces cerevisiae, the human dna ligase i, and with the vaccinia dna ligase. distant similari ...19921437556
biochemical construction of specific chimeric plasmids from cole1 dna and unfractionated escherichia coli dna.a series of chimeric plasmids was constructed using colicinigenic factor e1 (cole1) dna as the replicon and dna fragments carrying the galactose or tryptophan operons from e. coli. restriction endonuclease ecori digests of cole1 dna and various dnas containing the trp or gal operons were joined by t4 polynucleotide ligase [polynucleotide synthetase (atp), poly(deoxyribonucleotide):poly(deoxyribonucleotide) ligase (amp-forming), ec]. chimeric plasmids carrying the desired genes were selec ...1976792875
regulation of gene 32 expression during bacteriophage t4 infection of escherichia coli.the gene 32 protein of the bacteriophage t4 plays an important role in genetic recombination, dna repair, and dna replication; the protein functions in these processes by virtue of a strong binding capacity for single-stranded dna. during infections of escherichia coli by bacteriophage carrying amber of temperature-sensitive mutations in gene 32, the altered gene 32 protein (that is, the amber fragment of the missense polypeptide) is synthesized at greatly elevated rates. during infections by ph ...1976791947
effect of escherichia coli nusg function on lambda n-mediated transcription antitermination.the escherichia coli nus factors act in conjunction with the bacteriophage lambda n protein to suppress transcription termination on the lambda chromosome. nusa binds both n and rna polymerase and may also interact with other nus factors. to search for additional components of the n antitermination system, we isolated host revertants that restored n activity in nusa1 mutants. one revertant, nusg4, was mapped to the rif region of the e. coli chromosome and shown to represent a point mutation near ...19921531224
properties of condensed bacteriophage t4 dna isolated from escherichia coli infected with bacteriophage t4.methods developed for isolating bacterial nucleoids were applied to bacteria infected with phage t4. the replicating pool of t4 dna was isolated as a particle composed of condensed t4 dna and certain rna and protein components of the cell. the particles have a narrow sedimentation profile (weight-average s=2,500s) and have, on average, a t4 dna content similar to that of the infected cell. their dimensions observed via electron and fluorescence microscopy are similar to the dimensions of the int ...1976787557
construction and characterization of a chimeric plasmid composed of dna pfrom escherichia coli and drosophila melanogaster.a chimeric plasmid has been constructed in vitro from colicin e1 factor (col e1), nontransmissible r-factor rsf-1010, and drosophila melanogaster dnas by the sequential action of escherichia coli endonuclease ri(eco ri) and t4 phage dna ligase. the chimeric plasmid was assembled in two stages--first, a composite plasmid consisting of col e1 and rsf 1010 was constructed, followed by partial digestion of the composite with eco ri (in order to open one of the susceptible cleavage sites) and ligatio ...1975807234
heterologous deoxyribonucleic acid uptake and complexing with cellular constituents in competent bacillus subtilis.with competent cultures of bacillus subtilis the uptake of escherichia coli deoxyribonucleic acid (dna) is about 50% that for homologous dna. uptake of phage t6 dna, if any, is of the order of 7%, while nonglucosylated phage t6 (t6) dna is taken up almost as effectively as homologous dna. both t6 and t4 dna interfere only minimally with uptake of homologous dna; by contrast, t6 dna competes with homologous dna as effectively as the latter itself. these results indicate that the glucose residues ...1975811646
specificity of bacterial ribosomes and messenger ribonucleic acids in protein synthesis reactions in vitro.ribosomes from two gram-negative bacteria translated f2 rna, t4 early mrna, mrna from three gran-negative bacteria, and mrna from six gram-positive bacteria; ribosomes from three gram-positive bacteria translated mrna from the gram-positive strains, but did not translate the other mrnas. ribosomes from the gram-negative bacterium escherichia coli translated synthetic poly(u,g) but ribosomes from the gram-positive bacterium clostridium pasteurianum translated poly(u,g) very poorly, mrna from gram ...1976816792
immunoanalysis of ultraviolet radiation induced dna damage and repair within specific gene segments of plasmid dna.the region-specific heterogeneity of repairing dna damage has been established in several biological systems. a flexible and sensitive approach, based upon dna damage specific antibodies, is described to monitor the repair of specific lesions within discrete genomic segments. membrane transblotted dna restriction fragments are immunoanalyzed for the initial formation and repair of 254 nm radiation induced pyrimidine dimers. sensitivity of dimer immunodetection increases proportional to fragment ...19911657185
gene expression and stability of mrna affected by dna-arrested synthesis in gene 59, 46, and 47 mutants of bacteriophage t4.the effect of bacteriophage t4 gene 59 mutations (dna-arrested synthesis) on kinetics of dna synthesis, gene expression, and stability of mrna has been studied. when escherichia coli b was infected by a t4 gene 59 mutant, dna synthesis proceeded to increase linearly after initiation, but started to decrease at 8 min and was completely arrested at 12 min at 37 degrees c. at various incubation temperatures (20 to 42 degrees c), the initial rates and times of arrest of dna synthesis were different, ...1978702642
cleavage of nonglucosylated bacteriophage t4 deoxyribonucleic acid by restriction endonuclease eco ri.dnas lacking the glucosyl modification (glc-) and additionally lacking the 6-methylaminopurine (n6-methyladenine) modification (glc-, meade-) were prepared from appropriate t4 mutants. these dnas were cleaved by the purified restriction endonuclease eco ti from escherichia coli. normally modified dna (glc+, meade+) was not attached. the eco rii and the hemophilus enzymes hin dii and hin diii do not attack glc-, meade- t dna, possibly due to the presence of 6-hydroxymethylcytosine. eco ri produce ...19751090619
bacteriophage-host interaction and restriction of nonglucosylated t6.nonglucosylated t6 phage (t6gtam 16am30, hereafter called t6alpha gt-) were found to have two structural anomalies when compared with wild-type t6. the dna of t6alpha gt- phage contains single-strand interruptions. these can be seen both during infection, in the pool of replicating dna, and in dna extracted from purified phage. in addition, the sodium dodecyl sulfate-polyacrylamide gel pattern of t6alpha gt- phage structural proteins reveals a protein band not found in t6. the altered protein ha ...19751090750
characterization of new regulatory mutants of bacteriophage t4. ii. new class of mutants of bacteriophage t4 that overproduce the enzyme dihydrofolate reductase were investigated. unlike previously described overproducers of this enzyme (johnson and hall, 1974), these mutants did not overproduce deoxycytidylate deaminase. overproduction of dihydrofolate reductase by the new mutants occurred because enzymatic activity continued to increase for a longer period of time in cells infected by the mutants than in cells infected by wild-type phage. this continued increase occurr ...19751090753
oligonucleotide site directed mutagenesis of all histidine residues within the t4 endonuclease v gene: role in enzyme-nontarget dna order to evaluate the contributions that histidine residues might play both in the catalytic activities of endonuclease v and in binding to nontarget dna, the technique of oligonucleotide site directed mutagenesis was used to create mutations at each of the four histidine residues in the endonuclease v gene. although none of the histidines were shown to be absolutely required for the pyrimidine dimer specific dna glycosylase activity or the apurinic lyase activity, conservative amino acid cha ...19911868080
the human homologous pairing protein hpp-1 is specifically stimulated by the cognate single-stranded binding protein hrp-a.homologous pairing and strand exchange of dna are catalyzed by the human homologous pairing protein hpp-1 in a magnesium-dependent, atp-independent reaction that requires homologous dna substrates and stoichiometric quantities of hpp-1. here we show that the addition of the purified human single-strand binding (ssb) protein hrp-a to the reaction mixture stimulates the rate of homologous pairing 70-fold and reduces the amount of hpp-1 required for the reaction at least 10-fold. the identification ...19911924369
isolation and partial characterization of three escherichia coli mutants with altered transfer ribonucleic acid transfer ribonucleic acid (trna) methylase mutants were isolated from escherichia coli k-12 by examining the ability of rna prepared from clones of unselected mutagenized cells to accept methyl groups from s-adenosylmethionine catalyzed by crude enzymes from wild-type cells. five of the mutants had an altered uracil-trna methylase; consequently their trna's lacked ribothymidine. one mutant had trna deficient in 7-methylguanosine, and one mutant contained trna lacking 2-thio-5-methylaminome ...19751091626
transcription of azotobacter phage deoxyribonucleic acid. salt-dependent equilibrium between steps in initiation.the transcription of azotobacter phage a21 dna by escherichia coli or azotobacter vinelandii rna polymerase differs from that of some other dnas in its inhibition by moderate concentrations of kcl. this characteristic results in an apparent low template activity for this dna as compared with t4 dna under standard assay conditions. from an analysis of the dependence of the various steps in initiation on kcl it is concluded that the effect is exerted on an equilibrium between an inactive polymeras ...19751091643
bacteriophage t7 deoxyribonucleic acid replication in vitro. purification and properties of the gene 4 protein of bacteriophage t7.the t7 gene 4 protein, a protein known from genetic analysis to participate in phage dna replication in vivo, has been purified approximately 500-fold with an in vitro complementation assay. the protein, purified from cells infected with a t7 gene 4 temperature-sensitive mutant, is thermolabile, establishing that the complementation activity is in the protein product of the phage gene 4. the purified protein has no detectable nuclease, dna polymerase, or rna polymerase activity. however, in addi ...19751095580
direct selection of mutants restricting efficiency of suppression and misreading levels in e. coli b.we describe a method for the direct selection of e. coli mutants restricting efficiency of suppression and misreading levels using a t4-coded nonsense suppressor. one mutant isolated has the phenotype expected for a restrictive mutant and may be ribosomal. other possibilities are discussed.19751102920
reconstruction of bacteriophage t4 dna replication apparatus from purified components: rolling circle replication following de novo chain initiation on a single-stranded circular dna template.the protein products of t4 bacteriophage genes 41, 43, 45, 44, and 62 have been purified to near homogeneity using an assay which measures their stimulation of dna synthesis in a crude lysate of escherichia coli cells in fected by an appropriate mutant phage. when all of these proteins and t4 gene 32 protein are incubated in the presence of deoxyribonucleoside and ribonucleoside triphosphates, extensive dna synthesis occurs on both single and double-stranded dna templates. analysis of this in vi ...19751061070
recombination hotspots in bacteriophage t4 are dependent on replication origins.bacteriophage t4 recombination "hotspots" were first detected by the rescue of genetic markers from uv-irradiated phage particles. these hotspots have since been detected following treatments that yield other forms of dna damage, and at least one is active in the absence of damage. the previous mapping of phage replication origins near the peaks of two recombination hotspots suggested that the origins cause the localized enhancement of recombination. here we show that deletion of one origin elim ...19912068082
[the role of temperature as a factor of structure and functional fit of a "prey" virus based on the example of phage t4].by means of high-precision acoustic measurements and by the methods of fluorescent and electron microscopy investigations were performed of thermoinduced conformational changes in t4 bacteriophage and its thermolabile mutants altered in baseplate proteins (gene products "7", "8", "10"). a relationship was found between the conformational changes in t4 bacteriophage structure in the temperature range of 33-45 degrees c and the efficiency of bacteriophage adsorption and changes in the orientation ...19911809394
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