| signals in the phi 29 dna-terminal protein template for the initiation of phage phi 29 dna replication. | the protein-free terminal fragments hindiii b and l, from the left and right ends of phi 29 dna, respectively, but not internal fragments of similar size, were active as templates in the formation of the p3-damp initiation complex in an in vitro system containing purified phi 29 terminal protein p3 and dna polymerase p2, although the activity was lower than that obtained with the phi 29 dna-p3 complex. these results indicate the existence of specific sequences at the ends of phi 29 dna that allo ... | 1986 | 3097958 |
| synthesis of bacteriophage phi 29 proteins in bacillus subtilis. | seventeen bacteriophage phi29 proteins were detected in ultraviolet light-irradiated bacillus subtilis by autoradiography of polyacrylamide slab gels. the appearance of phi29 proteins occurred either before or concomitantly with viral dna replication. viral proteins detected early in the infectious cycle consisted of nine polypeptides ranging from 5,200 daltons to 54,000 daltons. two of the early proteins were identified as, respectively, the major capsid protein and the protein comprising the f ... | 1973 | 4199108 |
| proteins induced in bacillus subtilis infected with bacteriophage phi 29. | | 1973 | 4200881 |
| antigenic properties of bacteriophage phi 29 structural proteins. | serological methods and electron microscopy were used to study the structural proteins of the small bacillus subtilis bacteriophage phi29. this virus has a large number of fibers attached at both ends of its prolate head. a complex neck assembly is comprised of 12 symmetrically arranged appendages as the outer component. head fibers, neck appendages, and the head surface bind anti-phi29 antibodies. immune sera absorbed with defective lysates of suppressor-sensitive (sus) mutants have been used t ... | 1973 | 4202619 |
| bacteriophage t3 connector: three-dimensional structure and comparison with other viral head-tail connecting regions. | the bacteriophage t3 connector, which consists of 12 copies of protein gp8, has been studied by image processing of electron micrographs from negatively stained ordered aggregates. a three-dimensional reconstruction of t3 connectors was obtained by collection of tilted views and using the direct fourier method, up to 2.3 nm resolution. the reconstructed unit cell contains two connectors whose main structural features are essentially identical, but facing in opposite directions. the t3 connector ... | 1988 | 3262165 |
| a set of expression plasmids for the synthesis of fused and unfused polypeptides in escherichia coli. | a set of plasmid expression vectors for cloning of dna fragments containing open reading frames has been obtained. the plasmids carry the strong leftward promoter of bacteriophage lambda and the translation initiation signals from either the gene ner of bacteriophage mu or the gene 4 of bacteriophage phi 29. the vectors could overexpress the cloned sequences as fusion peptides at the n terminus with the n-terminal segment of the phi 29 protein p4 or at the c terminus with the escherichia coli be ... | 1987 | 3319785 |
| a precursor of the neck appendage protein of b. subtilis phage phi 29. | | 1974 | 4212893 |
| suppressor-sensitive mutants and genetic map of bacillus subtilis bacteriophage phi 29. | | 1974 | 4214301 |
| gene expression during the development of bacteriophage phi 29. 3. analysis of viral-specific protein synthesis with suppressible mutants. | fifty-four suppressible mutants of bacteriophage phi29 have been isolated with a variety of mutagens and assigned to eight complementation groups. viral-specific protein synthesis in uv light-irradiated, nonsuppressing bacillus subtilis 60084 was analyzed with exponential acrylamide gels. four additional phi29 proteins which were undetected on ordinary acrylamide gels are reported in this paper. five phage phi29 proteins have been unambiguously assigned to specific cistrons. two cistrons had ple ... | 1974 | 4362871 |
| the complete nucleotide sequence of the left very early region of escherichia coli bacteriophage prd1 coding for the terminal protein and the dna polymerase. | dna molecules replicating in a linear form have been found in certain viruses and plasmids of both prokaryotic and eukaryotic origin. characteristic of this type of molecules are the proteins covalently linked to their 5' ends and inverted terminal nucleotide sequences. the molecules replicate via a protein-priming mechanism, where participants include terminal protein and a specific polymerase. we report here the nucleotide sequence of the left very early region of escherichia coli bacteriophag ... | 1987 | 3322943 |
| transcription during the development of bacteriophage phi 29: production of host- and phi 29-specific ribonucleic acid. | the synthesis of ribonucleic acid (rna) during development of the virulent bacillus subtilis bacteriophage phi29 has been analyzed. transcription of host deoxyribonucleic acid (dna) continues at the preinfection rate throughout the latent period of viral growth. rna-dna hybridization was used to show that host messenger rna synthesis continues late into the phage lytic cycle. amino acid-labeling experiments show that this rna is continuously used to produce protein. ribosomal rna production is n ... | 1972 | 4630153 |
| characterization of the origins of replication of bacteriophage phi 29 dna. | the origins of replication of phi 29 dna have been studied by analyzing the activity as templates in the phi 29 in vitro replication system of e. coli recombinant plasmids and m13 derivatives containing phi 29 dna terminal sequences. plasmid pitr, containing the 6 bp long inverted terminal repeat of phi 29 dna, was shown to be essentially inactive. the analysis of a series of deletion derivatives of plasmid pid13, that contains the 73 and 269 bp from the left and right phi 29 dna ends, respectiv ... | 1988 | 3399382 |
| structure of bacillus subtilis bacteriophage phi 29 and the length of phi 29 deoxyribonucleic acid. | anderson, d. l. (university of minnesota, minneapolis), d. d. hickman, and b. e. reilly. structure of bacillus subtilis bacteriophage phi29 and the length of phi29 deoxyribonucleic acid. j. bacteriol. 91:2081-2089. 1966-bacillus subtilis bacteriophage phi29 were negatively stained with phosphotungstic acid. the head of phi29 has a hexagonal outline with a flattened base, and is about 315 a wide and 415 a in length. the virus has an intricate tail about 325 a in length. twelve spindle-shaped appe ... | 1966 | 4957028 |
| physical and biological properties of phage phi 29 deoxyribonucleic acid. | deoxyribonucleic acid (dna) molecules having a mean length of 5.8 mum were released from purified bacillus subtilis bacteriophage phi29 with 2 m sodium perchlorate. small 0.1 to 0.2-mum molecules were also detected in these dna preparations. since intact single chains annealed to form linear duplex molecules, phage phi29 dna was found to be nonpermuted. the molecular weights of single chains of phi29 dna were approximately half that of native dna, as determined by analytical band sedimentation i ... | 1968 | 4972802 |
| rescue of genetic markers from bacteriophage phi 29 dna fragments. | | 1970 | 4998256 |
| interaction of the bacteriophage phi 29 protein p6 with double-stranded dna. | the bacillus subtilis bacteriophage phi 29 protein p6 binds to double-stranded dna, but not to single-stranded dna, as determined by a gel retardation assay. the nature of the interaction was further studied by dnase i "footprinting" experiments. protein p6 binds to fragments containing the right or left terminal sequences of phi 29 dna, producing a characteristic pattern of hypersensitive bands spaced about 24 nucleotides apart along most of the fragment, flanking protected regions. binding of ... | 1988 | 3124105 |
| dna-protein complex in circular dna from phage phi-29. | | 1971 | 5002464 |
| mapping of temperature sensitive mutants of bacteriophage phi 29. | | 1972 | 5018452 |
| effect of the delta subunit of bacillus subtilis rna polymerase on initiation of rna synthesis at two bacteriophage phi 29 promoters. | initiation of rna synthesis by bacillus subtilis rna polymerase (sigma-43) has been examined at two early promoters of phage phi 29: the a2 promoter, which is a weak promoter, and the g2 promoter, which is a strong promoter. the delta subunit of the polymerase inhibits the rate of initiation at a2, but not g2. in addition, formation of stable complexes by the polymerase at a2, but not at g2, requires the presence of the first two nucleotides of the a2 transcript. | 1987 | 3126800 |
| temperature-sensitive mutants of bacteriophage phi-29. | | 1971 | 5167656 |
| effect of elevated temperature on deoxyribonucleic acid synthesis in bacteriophage phi-29-infected bacillus amyloliquefaciens. | deoxyribonucleic acid (dna) synthesis in bacteriophage phi29-infected bacillus amyloliquefaciens was studied at 37 and 45 c. infectious intracellular particles appear at the same time at both temperatures, but the average burst size is reduced 45 to 50% at 45 c. there is a transient inhibition of cellular mass increase at 45 c which is not observed at the lower temperature. in addition, the rate of host dna synthesis is reduced and the onset of viral-specific dna replication is delayed for 6 to ... | 1970 | 5497888 |
| initiation of phage phi 29 dna replication by mutants with deletions at the amino end of the terminal protein. | series of deletions at the amino end of protein p3, the phage phi 29 dna terminal protein (tp), have been constructed and characterized. measurements of the activity of the deletion mutants in the formation of the protein p3-damp initiation complex in vitro indicate the dispensability of the first 13 amino acids (aa) of the protein. the activity of protein p3 decreased considerably when 17 or more aa were deleted. the results on the in vitro phi 29 dna replication primed by the p3 deletion mutan ... | 1988 | 3133284 |
| restoration of direct fourier three-dimensional reconstructions of crystalline specimens by the method of convex projections. | we consider the problem of the three-dimensional (3-d) reconstruction of objects by the direct fourier method (dfm) and their restoration by the method of projections on to convex sets (pocs). the main discussion is centered on the case of specimens arranged in a two-dimensional (2-d) crystal and imaged by transmission electron microscopy, although the conclusions could be extended to more general cases. we present results of the restoration of the 3-d reconstruction of a computer generated 2-d ... | 1987 | 3573036 |
| inefficient translation of t7 late mrna by bacillus subtilis ribosomes. implications for species-specific translation. | bacillus subtilis 30 s subunits inefficiently recognize initiation sites in mrnas from gram-negative bacteria, but they are able to efficiently recognize initiation sites in mrna derived from gram-positive bacteria. mclaughlin et al. (mclaughlin, j. r., murray, c. l., and rabinowitz, j. c. (1981) j. biol. chem. 256, 11283-11291) have suggested that b. subtilis ribosomes require a strong shine-dalgarno sequence for translation initiation. to test whether this criterion is sufficient to explain th ... | 1985 | 3934154 |
| structure of phage phi 29 connector protein assembled in vivo. | the protein p10 that forms the connector of phage phi 29, has been produced in escherichia coli harboring a plasmid that carried the gene coding for this protein. the connector protein is assembled in a 13.4-s oligomer that has an apparent molecular weight of 460,000, suggesting that it is a dodecamer. the purified oligomers have been studied by electron microscopy of the isolated particles as well as by image-processing techniques (fourier and rotational filtering) of artificially induced two-d ... | 1985 | 3936270 |
| nucleotide sequence and transcription of a bacteriophage 29 early promoter. | we have studied the in vitro and in vivo transcription of a promoter for bacillus subtilis rna polymerase on bacteriophage phi 29 dna. the promoter is identified as an early promoter as it is transcribed in vitro by uninfected b. subtilis sigma 55-containing rna polymerase; is transcribed in vivo at both 7 min after infection and in the presence of chloramphenicol; and is transcribed right to left on the standard phi 29 map. the nucleotide sequence of the promoter and the initiation site for rna ... | 1985 | 3997806 |
| three-dimensional reconstruction of the connector of bacteriophage phi 29 at 1.8 nm resolution. | the three-dimensional reconstruction of the connector of bacteriophage phi 29 has been obtained from tilt series of negatively stained tetragonal ordered aggregates under low-dose conditions and up to a resolution of (1/1.8) nm-1. these connectors are built up as dodecamers of only one structural polypeptide (p10). two connectors form the crystal unit cell, each one facing in the opposite direction with respect to the plane of the crystal and partially overlapping. the main features of the two c ... | 1986 | 3586012 |
| three-dimensional reconstruction of bacteriophage phi 29 neck particles at 2 x 2 nm resolution. | the three-dimensional structure of the head-to-tail connecting region of bacteriophage phi 29 has been studied by analysing two-dimensional, hexagonal ordered aggregates of negatively stained viral necks to a resolution of 2 x 2 nm. these necks are composed of two proteins, p10 and p11; p10 being the connector protein. a 12-folded and a 6-folded axially symmetric domain are present in the specimen. the 12-folded domain is the larger part of the structure; it consists of 12 subunits associated in ... | 1985 | 4009722 |
| genetic study of suppressor-sensitive mutants of the bacillus subtilis bacteriophage phi 29. | with bacteriophage phi29 of bacillus subtilis 133, suppressor-sensitive (sus) hydroxylamine mutants have been isolated. intracistronic and intercistronic quantitative complementation placed the mutants in 13 cistrons, and three-factor crosses have been used to assign an unambiguous order for 10 cistrons. recombination frequencies have been presented for several regions of the genome to facilitate comparison of the sus system with the previously published temperature-sensitive mapping systems. | 1973 | 4196635 |
| characterization of a deletion mutant of bacteriophage phi 29. | a spontaneous deletion mutant of bacteriophage phi 29 (phi 29 delta 1) has been characterized. this mutant has a 1112-bp deletion, which covers almost the entire sequence of genes 14 and 15, including an early promoter (b2). while lysis is very delayed, the phage dna synthesis and internal phage development appear to be normal in the cells infected with this deletion mutant. these results indicate that the early functions are intact in phi 29 delta 1. our results also suggest that genes 14 and 1 ... | 1987 | 3590622 |
| in vitro initiation of bacteriophage phi 29 and m2 dna replication: genes required for formation of a complex between the terminal protein and 5'damp. | cell-free extracts prepared from phi 29 and m2-infected bacillus subtilis cells catalyse the formation of complexes between terminal protein and [alpha-32p]-damp in the presence of [alpha-32p]-datp, mgcl2, atp, and phage dna with terminal protein covalently linked at both the 5'ends. the complex formation does not take place when proteinase k-treated dna is added or when uninfected extract is used. the phi 29 complex thus formed is smaller than the m2 complex, primarily due to the different mole ... | 1983 | 6310350 |
| purification in an active form of the phage phi 29 protein p4 that controls the viral late transcription. | the phage phi 29 protein p4, that controls viral late transcription, was highly purified from escherichia coli cells harbouring a gene 4-containing plasmid. this protein, representing about 6% of the total cellular protein, was obtained in a highly purified form. the protein was characterized as p4 by amino acid analysis and nh2-terminal sequence determination. the purified protein was active in an in vitro transcription assay, allowing specific initiation of transcription at the phi 29 a3 late ... | 1987 | 3671066 |
| viral protein synthesis in bacteriophage phi 29-infected bacillus subtilis. | twenty-three (14)c-labeled phage phi29-specific proteins in lysates of uv-irradiated bacillus subtilis have been resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and identified by autoradiography. included in this group of proteins are the six major structural proteins of the virion. analysis of the temporal sequence of viral protein synthesis indicates that three groups of proteins can be identified by time of appearance, beginning at 2 to 4, 4 to 6, or 8 to 10 min after in ... | 1973 | 4203085 |
| analysis of bacteriophage phi 29 gene function: protein synthesis in suppressor-sensitive mutant infection of bacillus subtilis. | phage phi29 suppressor-sensitive (sus) mutants of 14 cistrons have been examined for production of (14)c-labeled viral-specific proteins in restrictive infections of bacillus subtilis. proteins specified by four cistrons (h, j, l, and n) have been resolved and identified by sodium dodecyl sulfate gel electrophoresis and autoradiography, and fragments of the normal polypeptides were detected. mutants of six cistrons (c, d, e, f, i, and m) demonstrated two or more missing bands in the gel profiles ... | 1974 | 4204249 |
| overproduction and purification of the connector protein of bacillus subtilis phage phi 29. | a phi 29 dna fragment containing genes 10 and 11, coding for the connector protein and the lower collar protein, respectively, has been cloned in the pbr322 derivative plasmid pkc30 under the control of the pl promoter of phage lambda. two polypeptides with the electrophoretic mobility of proteins p10 and p11 were labelled with 35s-methionine after heat induction. the proteins were characterized as p10 and p11 by radioimmunoassay and they represented about 10% and 7%, respectively, of the total ... | 1984 | 6324116 |
| computer graphic display method for visualizing three-dimensional biological structures. | a computer graphic display method that produces two-dimensional perspective views of three-dimensional objects is presented. the method is applied to the reconstruction at a resolution of 2.2 nanometers of the neck of bacteriophage phi 29, obtained from transmission electron micrographs processed by the direct fourier method. the combined use of directed illumination, reflectance models, color, and different levels of transparency provides a powerful tool for a better interpretation of the three ... | 1986 | 3754654 |
| transfecting deoxyribonucleic acid of bacillus bacteriophage phi 29 that is protease sensitive. | the transfecting activity of bacillus phage varphi29 dna, extracted either by sodium lauroyl sarcosine-phenol or by 2 m perchlorate, was destroyed by treatment with proteolytic enzymes, although these enzymes did not effect transfecting dnas of spp1, spo1, and sp50. these facts suggest that a protein is associated with transfective varphi29 dna. stabilization of protease-resistance during transfection appeared earlier than that of dnaseresistance, indicating that the protein associated with varp ... | 1972 | 4504368 |
| gene expression during the development of bacillus subtilis bacteriophage phi 29. i. analysis of viral-specific transcription by deoxyribonucleic acid-ribonucleic acid competition hybridization. | the ribonucleic acid (rna) specified by bacteriophage phi29 was analyzed to determine its composition at various times in the viral lytic cycle. although viral-specific rna was detected immediately after infection, a large increase in the rate was observed at 10 min when dna synthesis began. phi29 was found to resemble other viruses in that gene expression occurred in two stages which could be defined temporally as "early" and "late." early rna appeared before the onset of viral deoxyribonucleic ... | 1973 | 4630802 |
| factors involved in the initiation of phage phi 29 dna replication in vitro: requirement of the gene 2 product for the formation of the protein p3-damp complex. | to study the requirements for the in vitro formation of the protein p3-damp complex, the first step in phi29 dna replication, extracts from b. subtilis infected with phi29 mutants in genes 2, 3, 5, 6 and 17, involved in dna synthesis, have been used. the formation of the initiation complex is completely dependent on the presence of a functional gene 2 product, in addition to protein p3 and phi29 dna-protein p3 as template. atp is also required, although it can be replaced by other nucleotides. t ... | 1983 | 6402761 |
| nucleotide sequence of phage phi 29 gene 7: structure of intergenic spacer between the major early and late genes. | | 1986 | 3763399 |
| nucleotide sequence of the late region of bacillus phage phi 29 completes the 19,285-bp sequence of phi 29 genome. comparison with the homologous sequence of phage pza. | the 12,177-bp nucleotide (nt) sequence of the late region of bacillus phage luminal diameter 29 genome was determined. this sequence completes the entire 19,285-bp sequence of phage luminal diameter 29 dna. eleven open reading frames were found in this region, and these were assigned to eleven late genes. ribosome-binding sites and a potential transcriptional promoter and terminator are considered. the nt sequence was compared to the homologous region of the closely related phage pza and tolerat ... | 1986 | 3803926 |
| replication of phage phi 29 dna with purified terminal protein and dna polymerase: synthesis of full-length phi 29 dna. | a system that replicates bacteriophage phi 29 dna with protein p3 covalently attached to the two 5' ends, using as the only proteins the phi 29 dna polymerase and the terminal protein, is described. restriction analysis of the 32p-labeled dna synthesized in vitro showed that all phi 29 dna fragments were labeled. analysis by alkaline sucrose gradient centrifugation of the dna labeled during a 10-min pulse showed that, after a 20-min chase, about half of the dna molecules had reached apparently f ... | 1985 | 3863101 |
| a novel dna polymerase induced by bacillus subtilis phage phi 29. | a novel dna polymerase induced by bacillus subtilis bacteriophage phi 29 has been identified. this polymerase can be separated from host dna polymerase, by fractionation of extracts prepared from phage infected cells, using phosphocellulose chromatography. the isolated polymerase prefers poly(da)oligo(dt) as template. the dna polymerase isolated from the cells infected with a gene 2 temperature sensitive mutant (ts2) showed greater heat-lability than that induced by wild type phi 29. the ts2 dna ... | 1983 | 6424095 |
| morphogenesis of bacteriophage phi 29 of bacillus subtilis: prohead restoration for dna-gp3 packaging and assembly. | the dna-protein complex dna-gp3 of phi 29 is efficiently packaged into purified proheads with the aid of plasmid-derived gp16. the filled heads can be assembled to phage by addition of an extract providing the products for neck-tail assembly (bjornsti et al., j. virol. 50:766-772, 1984). however, purified proheads lost their competence to package dna-gp3 upon storage for 2 months at 4 degrees c. competence was restored by complementation with extracts of certain mutant-infected cells, and these ... | 1985 | 3919187 |
| nucleotide sequence of the major early region of bacillus subtilis phage pza, a close relative of phi 29. | the 5200-bp nucleotide sequence of the major early region of bacteriophage pza has been determined. open reading frames (orfs) and potential transcriptional and translational regulatory signals were found in this region. the sequence was compared with the known sequence of the homologous region of the closely related phage phi 29 (yoshikawa and ito, 1982). this comparison permitted a more accurate assignment of several orfs and regulatory signals. | 1985 | 3934048 |
| a genetic study of temperature-sensitive mutants of the bacillus subtilis bacteriophage phi 29. | | 1971 | 5000908 |
| a defined system for in vitro packaging of dna-gp3 of the bacillus subtilis bacteriophage phi 29. | the bacteriophage phi 29 dna-gene product 3 complex (dna-gp3) has been efficiently packaged into proheads in a completely defined in vitro system. the phi 29 dna packaging protein gp16, the product of gene 16, was overproduced in escherichia coli and purified to near homogeneity. the purified gp16 packaged 23% of the dna-gp3 added to purified proheads in the defined mixture, while gp16 in an extract of phage-infected cells packaged 26% of the dna-gp3. no host proteins were required in the define ... | 1986 | 3458193 |
| temperature-shift analysis of bacteriophage phi 29 gene expression in bacillus amyloliquefaciens. | temperature shift-up experiments with conditional lethal mutants of bacillus phage phi29 have allowed placement of early, middle, and late functions on the linear phi29 genetic map most of the phi29 cistrons are late and are found at the ends of the map. | 1971 | 5119489 |
| cloning and purification of a unique lysozyme produced by bacillus phage phi 29. | a dna fragment of the bacteriophage phi 29 chromosome, encoding the entire sequence of phi 29 gene 15, has been cloned into the escherichia coli expression vector pplc245 under the control of the phage lambda major leftward promoter, pl. upon heat induction, a protein with an apparent molecular mass of 26 kda was overproduced. the molecular mass of this protein corresponds to the 28 kda predicted for the product of gene 15 from its nucleotide sequence. the overproduced protein has been purified ... | 1987 | 3469652 |
| morphogenesis of bacteriophage phi 29 of bacillus subtilis: oriented and quantized in vitro packaging of dna protein gp3. | the assembly of phage phi 29 occurs by a single pathway, and the dna protein (dna-gp3) of "packaging intermediates" can be obtained after dnase i interruption of in vitro complementation. a broad spectrum of dna molecules of variable length was isolated from dnase i-treated proheads. restriction endonuclease ecori digestion and electrophoretic analysis of these dna molecules suggested that dna-gp3 packaging was oriented with respect to the physical map and was a complex process. proteinase k-tre ... | 1983 | 6185695 |
| specificity of promoter site utilization in vitro by bacterial rna polymerases on bacillus phage phi 29 dna. transcription mapping with exonuclease iii. | bacillus subtilis rna polymerase holoenzyme transcribes phi 29 dna in vitro producing five major rna species defined by characteristic electrophoretic mobilities. in addition to these products, escherichia coli rna polymerase transcribes phi 29 dna to yield three rna species not detected when transcribing with the b. subtilis enzyme under the same optimal reaction conditions for rna synthesis. transcriptional analysis of purified restriction fragments and exonuclease iii-digested dna established ... | 1980 | 6251067 |
| the protein covalently linked to the 5' termini of the dna of bacillus subtilis phage phi 29 is involved in the initiation of dna replication. | | 1980 | 6771916 |
| nucleotide sequence at the termini of the dna of bacillus subtilis phage phi 29. | phage phi 29 dna cannot be phosphorylated with polynucleotide kinase and [gamma-32p]atp because of the presence of a viral protein covalently linked to the 5' termini. the 5' ends can, however, be made susceptible to phosphorylation by treatment with alkali and alkaline phosphatase. restriction fragments hpa ii c and hpa ii f, corresponding to the right and left ends of phi 29 dna, respectively, were labeled at the 5' ends with polynucleotide kinase and [gamma-32p]atp or at the 3' ends with term ... | 1981 | 6262800 |
| nucleotide sequences of transcription and translation initiation regions in bacillus phage phi 29 early genes. | phi 29 dna directs the synthesis of three major proteins of mr = 22,400, 13,900, and 10,500 in a cell-free transcription-translation system derived from bacillus subtilis. we have determined the locations of the coding regions for these early proteins on the phi 29 genome, and our results are in agreement with genetic evidence that the 22.4-kilodalton protein is the product of cistron 17 (p17) and the 13.9-kilodalton protein is the product of cistron 6 (p6). the 13.9-kilodalton and 10.5-kilodalt ... | 1982 | 6274853 |
| purification, properties and assembly of the neck-appendage protein of the bacillus subtilis phage phi 29. | the purification of the neck appendage protein of phi 29, p12*, which is involved in the adsorption of the phage to bacillus subtilis, is described. the purified native protein is in a dimeric form and can be assembled, in vitro, onto purified 12- particles that lack the neck appendages, suggesting that the incorporation of p12* to the rest of the phage structure is a self-assembly process. the assembly of protein p12* in vitro follows cooperative kinetics and it occurs with an efficiency of abo ... | 1981 | 6793359 |
| high level synthesis in escherichia coli of the bacillus subtilis phage phi 29 proteins p3 and p4 under the control of phage lambda pl promoter. | the hind iii g fragment from the bacillus subtilis phage phi 29 dna, inserted downstream from the bacteriophage lambda promoter pl carried by a pbr322 derivative plasmid (pplc28), directed the synthesis in e. coli of two proteins of apparent molecular weight 27500 and 12500. with the use of the recombinants obtained with the dna from mutants sus3(91) and sus4(56), the two proteins were identified as a modified p3 (p3'), the protein covalently linked to the 5' ends of phi 29 dna, and p4, responsi ... | 1982 | 6292851 |
| nucleotide sequence of the early genes 3 and 4 of bacteriophage phi 29. | the nucleotide sequence of an early region of the phi 29 genome has been determined. the sequenced region includes genes 3 and 4, which code for the protein covalently linked to the 5' ends of phi 29 dna and the protein involved in the control of late transcription, respectively. the position and nature of the mutations of mutants sus3(91) and sus4(56) has also been determined. | 1982 | 6292852 |
| cloning and expression in escherichia coli of the gene coding for the protein linked to the ends of bacillus subtilis phage phi 29 dna. | a phi 29 dna fragment containing gene 3, coding for the 5'-terminal protein, and several other early genes has been cloned in a pbr322 derivative plasmid (pkc30) under the control of the pl promoter of bacteriophage lambda. four polypeptides of mr 27000, 18500, 17500 and 12500 were labelled with [35s]methionine after heat induction, accounting for about 15% of the de novo synthesized protein. the mr 27000 and 12500 proteins were characterized as p3, the 5'-terminal protein, and p4, involved in t ... | 1983 | 6301951 |
| structure of the head-tail connector of bacteriophage phi 29. | | 1982 | 6804634 |
| initiation of phage phi 29 dna replication by the terminal protein modified at the carboxyl end. | a mutant at the carboxyl end of the terminal protein, p3, of phage phi 29 dna has been constructed by inserting an containing the stop translation codon tga in the three possible reading frames, immediately downstream of a phage phi 29 dna fragment coding for all but the last five amino acids of protein p3. the activity in the formation of the p3-damp initiation complex in vitro of this mutant as well as another one previously isolated, also mutated at the carboxyl end, have been tested. the re ... | 1983 | 6316260 |
| template requirements for initiation of phage phi 29 dna replication in vitro. | the template requirements for the formation of the phi 29 protein p3-damp initiation complex in vitro have been studied. the initiation reaction requires the parental protein p3 but not an intact dna molecule. protein p3-containing fragments from the left- or right-hand dna ends were active as template for formation of the initiation complex provided they had a minimal size: a 26-base-pair-long fragment was active whereas a 10-base-pair-long one was essentially inactive. however, the activity of ... | 1984 | 6320176 |
| in vitro transcription of the bacillus subtilis phage phi 29 dna by bacillus subtilis and escherichia coli rna polymerases. | the escherichia coli rna polymerase bound to phage phi 29 dna has been visualized by electron microscopy. thirteen specific binding sites have been observed at 1.7,2.6,5.5,10.4,13.7,25.2,25.7,26.3,33.5,59.5,69.2,91.7 and 99.6 dna length units and they have been named a1,a1i,a1ii,a1iii,a1iv,a2,a2i, a3, a4,b1,b1i,c1 and c2, respectively. the binding sites a1,a2,a3,b1,c1 and c2 coincide with those found with bacillus subtilis rna polymerase. the transcription of phage phi 29 dna with b. subtilis or ... | 1984 | 6322128 |
| overproduction and purification of the gene 2 product involved in the initiation of phage phi 29 replication. | | 1984 | 6395657 |
| in vitro replication of bacteriophage phi 29. | | 1984 | 6395662 |
| structural localization of the proteins of the head to tail connecting region of bacteriophage phi 29. | the head to tail connector of bacteriophage phi 29 has been studied to locate its two structural proteins (p10 and p11). treatment with trypsin led to proteolysis of p10 while p11 remained intact. computer filtration of electron micrographs of crystals of trypsinized necks showed a change in the external 12-fold area of the neck when compared with control necks. proteolized necks completely released p10 after treatment with low concentrations of an ionic detergent. the resulting structures, cont ... | 1983 | 6401885 |
| assembly of the tail protein of the bacillus subtilis phage phi 29. | by in vitro complementation we have determined that gene 13 product functions during phi 29 morphogenesis after the formation of 11- particles, specifically in the functional assembly of the tail protein, p9. protein p9 from 8- but not from 8-13- extracts assembles in vitro into either 11-13- or 12-13- particles. the action of gene 13 product on p9 for its correct assembly has to take place in vivo; no complementation of 12-13- and 9- lysates occurs in vitro. protein p9, isolated from phi 29-inf ... | 1983 | 6402855 |
| in vitro synthesis of late bacteriophage phi 29 rna. | a crude p-100 fraction prepared from bacillus subtilis 21 min after infection with wild-type phage phi 29 supported the in vitro synthesis of late phi 29 rna by added rna polymerase. synthesis of late rna was also detected when purified phi 29 dna was transcribed by rna polymerase in the presence of an s-150 fraction obtained by lysis of phi 29-infected cells in the presence of 1 m nacl. late phi 29 rna was not synthesized when either the p-100 or the s-150 fraction was prepared from cultures in ... | 1983 | 6406684 |
| protein-primed initiation of phage phi 29 dna replication. | we recently reported the development of an in vitro replication system for bacteriophage phi 29 dna. we have used this system for the isolation of replication activity associated with gene 3 protein (terminal protein) from phi 29-infected bacillus subtilis cells. we utilized two assay systems: (i) dna replication dependent on phi 29 dna with the 5' end covalently linked to terminal protein (dna-protein) and (ii) the formation of complex between the terminal protein and damp. the dna-replication ... | 1983 | 6410387 |
| purification in a functional form of the terminal protein of bacillus subtilis phage phi 29. | phage phi 29 terminal protein, p3, essentially pure, was isolated in a denatured form from viral particles, and anti-p3 antiserum was obtained. a radioimmunoassay to detect and quantitate protein p3 was developed. by using this assay, native protein p3 was highly purified from escherichia coli cells harboring a gene 3-containing recombinant plasmid. after three purification steps, the protein was more than 96% pure; its amino acid composition was very similar to that deduced from the nucleotide ... | 1984 | 6424120 |
| circularly permuted viral prna active and specific in the packaging of bacteriophage phi 29 dna. | a viral-encoded 120-base prna has been shown to have an essential role in the packaging of bacteriophage phi 29 dna. the finding that both the 5'- and 3'-termini of the prna are proximate and crucial for biological function (c. zhang, c. lee, and p. guo, 1994, virology, 201, 77-85) prompted investigation of the activity of circularly permuted prnas (cprna) and of the expandability and essentiality of bases extending from the termini. a 117-base prna with a deletion of three bases downstream of t ... | 1995 | 7533964 |
| structure of bacteriophage phi 29 dna. | | 1984 | 6427030 |
| bacteriophage phi 29 proteins required for in vitro dna-gp3 packaging. | in vitro assembly of bacteriophage phi 29 in crude extracts involves efficient packaging of a dna-protein complex (dna- gp3 ) into a prohead with the aid of the gene 16 product ( gp16 ) and subsequent assembly of neck and tail proteins ( bjornsti et al., j. virol. 41:508-517, 1982; bjornsti et al., j. virol. 45:383-396, 1983; bjornsti et al., proc. natl. acad. sci. u.s.a. 78:5861-5865, 1981). to define the viral proteins required for the dna- gp3 encapsidation phase, we purified biologically act ... | 1984 | 6427474 |
| cloning and expression of gene 2, required for the protein-primed initiation of the bacillus subtilis phage phi 29 dna replication. | a phi 29 dna fragment containing gene 2, coding for a phi 29-specific dna polymerase required for the formation of the terminal protein p3-damp initiation complex, the first step in phi 29 dna replication, has been cloned in plasmid pplc28 under the control of the pl promoter of bacteriophage lambda. four polypeptides of mr 68 000, 5800 and 3400 and less than 2000 were labelled with [35s]methionine after heat induction. the protein of mr 68 000 had the size expected for protein p2 and it account ... | 1984 | 6092229 |
| cloning, nucleotide sequence and high level expression of the gene coding for the connector protein of bacillus subtilis phage phi 29. | the phi 29 dna restriction fragment hindiii-d, shown to contain gene 10 coding for the connector protein, has been cloned in plasmid pplc28 under the control of the pl promoter of phage lambda. after heat induction to inactivate the lambda repressor, a protein with the electrophoretic mobility of the connector protein p10 was synthesized, accounting for about 30% of the total escherichia coli protein after 3 h of induction. the 2205 nucleotide-long sequence of the cloned hindiii-d fragment has b ... | 1984 | 6096227 |
| structure of replicating dna molecules of bacillus subtilis bacteriophage phi 29. | we isolated phi 29 dna replicative intermediates from extracts of phage-infected bacillus subtilis, pulsed-labeled with [3h]thymidine, by velocity sedimentation in neutral sucrose followed by cscl equilibrium density gradient centrifugation. during a chase, the dna with a higher sedimentation coefficient in neutral sucrose and a lower sedimentation rate in alkaline sucrose than that of viral phi 29 dna was converted into mature dna. the material with a density higher than that of mature phi 29 d ... | 1980 | 6768899 |
| protein p3 is linked to the dna of phage phi 29 through a phosphoester bond between serine and 5'-damp. | to investigate the role of protein p3 in bacteriophage phi 29 initiation of replication, we have studied the nature of the covalent linkage between protein p3 and phi 29 dna. the protein-dna compound was digested with micrococcal nuclease and pronase resulting in a nucleotidyl-peptide that was further digested by alkaline phosphatase and snake venom phosphodiesterase yielding 5'-damp. the dna-protein linkage is sensitive to alkali. treatment of the nucleotidyl-peptide with 0.1 m naoh at 37 degre ... | 1980 | 6779279 |
| bacteriophage phi 29 infection of bacillus subtilis minicells. | bacteriophage phi 29 can infect b. subtilis minicells and synthesize all the phage-coded proteins detected in ultraviolet irradiated-infected b. subtilis cells. synthesis of phage unit-length dna has been obtained after infection of minicells with phi 29. the dna can be encapsulated in particles with a sedimentation rate similar to that of phage phi 29 produced in b. subtilis cells. the particles produced in minicells can be adsorbed to b. subtilis cells, but infectivity has not been demonstrate ... | 1980 | 6780760 |
| initiation factor-independent translation of mrnas from gram-positive bacteria. | initiation factor-independent translation of mrna derived from bacillus phage phi29 dna occurs with translation systems derived from bacillus subtilis or escherichia coli. this is in sharp contrast to the strict dependence on ribosome salt wash fraction of e. coli ribosomes for the translation of t7 and other mrnas derived from gram-negative organisms. | 1981 | 6795625 |
| dual translational start motif evolutionarily conserved in the holin gene of bacillus subtilis phage phi 29. | holins represent phage encoded lysis functions required for transit of the phage murein hydrolases to the periplasm. the lambda s, phage 21 s, and p22 13 holin genes contain a dual translational start motif, beginning with met1-lys2-x-met3. in all cases both start codons at the 5' end of the respective holin gene are utilized. the resulting polypeptides have opposing functions, with the longer product acting as an inhibitor of the shorter one. the 131-codon gene 14 of bacillus subtilis phage phi ... | 1995 | 7831803 |
| in vitro assembly of the bacillus subtilis bacteriophage phi 29. | in vitro assembly of the bacillus subtilis bacteriophage phi 29 that approaches the efficiency of assembly in vivo has been demonstrated. proheads, dna, and gene 16 product (gp16) were essential for dna encapsidation, and the average yield in extracts was 180 phage per prohead donor cell. the in vitro maturation was very similar to in vivo assembly in terms of yield, intermediates, and abortive structures. more that 30% of the proheads in the extract were converted to phage, and about 20% of dna ... | 1981 | 6795639 |
| structure of protein-containing replicative intermediates of bacillus subtilis phage phi 29 dna. | | 1982 | 6801848 |
| cloning, molecular characterization, and expression of the genes encoding the lytic functions of lactococcal bacteriophage phi lc3: a dual lysis system of modular design. | the genes encoding the lysis proteins of lactococcus lactis bacteriophage phi lc3 were cloned, sequenced, and expressed in escherichia coli. the phi lc3 lysis genes, lysa and lysb, encode a membrane-disrupting protein (lysa) of 88 amino acids, and a cell wall degrading protein (lysb) of 429 amino acids, which shares significant sequence similarity with lysins from the streptococcus pneumoniae phages cp-1, cp-7, and cp-9, and lactobacillus delbrueckii phage mv1. both lysa and lysb function in e. ... | 1994 | 7922887 |
| morphogenesis of bacteriophage phi 29 of bacillus subtilis: dna-gp3 intermediate in in vivo and in vitro assembly. | the assembly of phage phi 29 occurs by a single pathway, and dna-protein (dna-gp3) has been shown to be an intermediate on the assembly pathway by a highly efficient in vitro complementation. at 30 degrees c, about one-half of the viral dna synthesized was assembled into mature phage, and the absolute plating efficiency of phi 29 approached unity. dna packaging at 45 degrees c was comparable to that at 30 degrees c, but the burst size was reduced by one-third. when cells infected with mutant ts3 ... | 1982 | 6804642 |
| rna polymerase of myxococcus xanthus: purification and selective transcription in vitro with bacteriophage templates. | dna-dependent rna polymerase from vegetative cells of the gram-negative, fruiting bacterium myxococcus xanthus was purified more than 300-fold by a modified burgess procedure (lowe et al., biochemistry 18:1344-1352, 1979), using polymin p precipitation, 40 to 65% saturated ammonium sulfate fractional precipitation, double-stranded dna cellulose chromatography, a5m gel filtration chromatography, and single-stranded dna agarose chromatography. the last step separated the rna polymerase into a core ... | 1982 | 6806251 |
| in vitro complex formation between bacteriophage phi 29 terminal protein and deoxynucleotide. | | 1982 | 6807309 |
| nucleotide sequence of the major early region of bacteriophage phi 29. | the nucleotide sequence of the left end of bacteriophage phi 29 dna has been determined. together with data reported earlier (yoshikawa et al., 1981), this sequencing comprises the major early genetic region of this viral genome (5708 bp). computer analysis of the dna sequences revealed that there are up to fifteen open reading frames which could encode polypeptides containing more than thirty amino acids. the dna sequence also revealed a number of potential regulatory signals, promoters and rib ... | 1982 | 6809534 |
| in vitro replication of bacteriophage phi 29 dna. | we have been studying the mechanisms of linear dna replication by using bacillus bacteriophage phi 29 as a model system. to isolate and characterize the proteins required for phi 29 dna replication, we have developed a cell-free replication system. a cell-free extract prepared from phi 29-infected bacillus subtilis catalyzes the semiconservative replication of phi 29 dna, but only if exogenous phi 29 dna-protein complex is used as the template. this template consists of linear duplex dna with a ... | 1982 | 6813856 |
| a highly sensitive system for the in vitro assembly of bacteriophage phi 29 of bacillus subtilis. | a sensitive system for the assay of bacteriophage phi 29 assembly in vitro was developed using 12 recombinant proteins and synthetic prna. this system detected in vitro assembled infectious phages up to 10(7) plaque forming units (pfu) per milliliter without any background. phi 29 dna-gp3 concentration dependence in phage assembly was found to be first order, while the dna-packaging protein gp16 dependence was higher order. the requirement for specific phi 29 prna for phi 29 dna packaging was co ... | 1994 | 8030206 |
| initiation of phage phi 29 dna replication in vitro: formation of a covalent complex between the terminal protein, p3, and 5'-damp. | incubation of extracts of phi 29-infected bacillus subtilis with [alpha-32p]datp produced a labeled protein having the electrophoretic mobility of p3, the 5'-terminal protein of phi 29 dna. the reaction product was resistant to treatment with micrococcal nuclease, phosphatase, and rnases a and t1 and sensitive to proteinase k. incubation of the 32p-labeled protein with piperidine under conditions in which the phi 29 dna-protein p3 linkage is hydrolyzed released 5'-damp. the reaction with [alpha- ... | 1982 | 6813861 |
| terminal proteins and short inverted terminal repeats of the small bacillus bacteriophage genomes. | the genome of bacillus phage phi 29 contains covalently linked protein at both ends. these dna terminal proteins are essential for phi 29 dna replication. we have isolated phi 29 terminal protein from each end separately and compared their two-dimensional peptide maps. our results showed the two proteins to be identical. the dnas of four phages examined (phi 15, nf, m2y, and ga-1) also contain protein at both ends of the dna molecules. the chymotryptic peptide maps of these dna terminal proteins ... | 1981 | 6941313 |
| adsorption of bacteriophage phi 29 to bacillus subtilis through the neck appendages of the viral particle. | phage phi 29 particles produced under restrictive conditions by mutants in gene 12 have normal amounts of all of the structural proteins except the appendage protein, p12*, which is missing. these particles are not infective and do not adsorb to bacillus subtilis cells. by in vitro complementation of 12- particles with extracts containing protein p12* or with purified protein p12*, the defective particles could bind the appendage protein and become infective and able to adsorb to bacteria. there ... | 1981 | 7241648 |
| dna replication of bacteriophage phi 29: characterization of the intermediates and location of the termini of replication. | | 1980 | 7395108 |
| helix-destabilizing activity of phi 29 single-stranded dna binding protein: effect on the elongation rate during strand displacement dna replication. | the single-stranded dna (ssdna)-binding protein (ssb) of bacteriophage phi 29 is one of the virus-encoded proteins required for viral dna replication. we have found that phi 29 ssb has helix-destabilizing activity since it removes secondary structure of the ssdna in phi 29 replicative intermediates, as revealed by electron microscopy, and displaces oligonucleotides annealed to m13 ssdna. to investigate the mechanism of the ssb-dependent stimulation of phi 29 dna replication we have characterized ... | 1995 | 7473731 |
| in vitro assembly of infectious virions of double-stranded dna phage phi 29 from cloned gene products and synthetic nucleic acids. | up to 6 x 10(7) pfu of infectious virions of the double-stranded dna bacteriophage phi 29 per ml were assembled in vitro, with 11 proteins derived from cloned genes and nucleic acids synthesized separately. the genomic dna-gp3 protein conjugate was efficiently packaged into a purified recombinant procapsid with the aid of a small viral rna (prna) transcript, a dna-packaging atpase (gp16), and atp. the dna-filled capsids were subsequently converted into infectious virions after the addition of fo ... | 1995 | 7609071 |
| characterization and purification of a phage phi 29-encoded dna polymerase required for the initiation of replication. | the phage phi 29 protein p2, required for the formation of the protein p3-damp initiation complex, has been purified from escherichia coli cells harboring a gene 2-containing recombinant plasmid. the purified protein p2, of molecular weight 68,000, had a specific dna polymerase activity that elongated the p3-damp initiation complex when phi 29 dna-protein p3 was used as template. in addition, the purified protein p2 was active in catalyzing the initiation reaction when complemented with phi 29 m ... | 1984 | 6433348 |
| replication of bacteriophage phi 29 dna in vitro: the roles of terminal protein and dna polymerase. | phi 29 dna replication is initiated by the formation of a covalent complex between the viral-coded terminal protein and damp (tp-damp). this initiation reaction system has been reconstituted from two phage-encoded proteins, the terminal protein and dna polymerase. the phi 29 dna polymerase was purified from phage-infected cells by using poly(da) x p(dt)12-18 as an assay template. the purified polymerase has an apparent molecular mass of 68 kda in its native form and it appears to function as a m ... | 1984 | 6433349 |
| bacteriophage phi 29 dna replication in vitro: participation of the terminal protein and the gene 2 product in elongation. | from phi 29-infected bacillus subtilis cells, we have isolated a protein fraction which promotes in vitro replication of phi 29 dna. this fraction catalyses both initiation and elongation, indicating that it contains the product of gene 3 (tp: terminal protein) and the product of gene 2 (gp2: probably a dna polymerase), since initiation requires the two products (blanco et al. 1983; matsumoto et al. 1983). the fractions isolated from cells infected with temperature-sensitive (ts) mutants of gene ... | 1984 | 6438445 |
| structural study of tetragonal-ordered aggregates of phage phi 29 necks. | a new class of two-dimensional tetragonal aggregates of phage phi 29 necks has been studied by electron microscopy and a combination of fourier filtering procedures and detailed rotational analysis. the results confirm the main features of the head-to-tail connecting region previously observed in hexagonal aggregates. there are several differences in the resulting pictures that can be attributed to the different way in which the aggregates are organized and stained. | 1984 | 6544883 |
| characterization of an rna-binding domain in the bacteriophage phi 29 connector. | the connector of bacteriophage phi 29 is known to promote the viral prohead assembly, to bind dna, and to drive dna packaging into preformed viral shells in an rna-dependent process. in this report, the phi 29 connector protein, p10, is shown to bind rna in a sequence-independent fashion, and to possess an rna recognition motif comprised approximately the region between residues 21 and 94 of the p10 sequence. substitution mutants in specific amino acids of the rna-binding domain obtained by site ... | 1993 | 7690751 |
| phage phi 29 protein p6: a viral histone-like protein. | phage phi 29 protein p6 is one of the most abundant viral proteins in phi 29-infected b subtilis cells, constituting about 4% of the total cellular proteins (about 3 x 10(6) copies/cell) at late infection. electron microscopic studies showed that, in vitro, protein p6 forms heterogeneously-sized complexes all along phi 29 dna, suggesting that protein p6 may have a role in genome packaging and organization. the low stability of the protein p6-phi 29 dna complexes observed in vitro could reflect t ... | 1994 | 7748942 |