nucleotide sequence of the cohesive single-stranded ends of bacillus subtilis temperate bacteriophage phi 105.the cohesive single-stranded ends of temperate bacillus subtilis phage phi 105 were analyzed with the exonuclease activities of the klenow fragment of dna polymerase i and with exonuclease iii and were found to be 3' extensions. chemical sequencing of 3'-end-labeled fragments showed that the ends are 7-base extended 3' single strands and have the sequence: 5'-gcgctcc-3'. 3'-cgcgagg-5'19853927011
construction of improved bacteriophage phi 105 vectors for cloning by transfection in bacillus subtilis.a series of improved phage vectors have been constructed, based on bacillus subtilis bacteriophage phi 105, which can be used to clone genes in b. subtilis by direct transfection of protoplasts. the new vectors, designated phi 105j23, phi 105j24, phi 105j27 and phi 105j28, show frequencies of plaque formation that are equal to those of wild-type phi 105. this represents at least a 10-fold improvement over phi 105j9, the vector used in previous cloning experiments. two of the new vectors phi 105j ...19873116161
cloning of sporulation gene spoivc in bacillus subtilis.sporulation gene spoivc of bacillus subtilis was cloned by the prophage transformation method in temperate phage phi 105. the specialized transducing phage, phi 105 spoivc-1, restored the sporulation of the asporogenous mutant of b. subtilis strain 1s47 (spoivc133). transformation experiments showed that the spoivc gene resides on a 7.3 kb hindiii restriction fragment. subsequent analysis of the 7.3 kb hindiii fragment with restriction endonuclease ecori showed that the spoivc gene resides on a ...19852993797
nucleotide sequence of the immunity region of bacillus subtilis bacteriophage phi 105: identification of the repressor gene and its mrna and protein products.a gene involved in the regulation of lysogeny in the temperate bacillus subtilis phage phi 105 has been identified and isolated. a plasmid, pdc4, was constructed that contains a 740-bp hindiii-pvuii fragment that is derived from the phi 105 immunity region and is capable of rendering b. subtilis immune to infection by phi 105. three different hybrid plasmids that contain the 740-bp fragment, pag101 [cully and garro, j. virol. 34 (1980) 789-791], pdc1 and pdc2, were found to synthesize a common 1 ...19853934047
heat induction of prophage phi 105 in bacillus subtilis: bacteriophage-induced bidirectional replication of the bacterial chromosome.a mutant of bacillus subtilis, dna-1, which cannot initiate new rounds of dna replication (obtained from n. sueoka) was lysogenized with wild-type phi 105 and with the heat-inducible mutant phi 105 cts23. bacteria were incubated at the permissive temperature in the presence of chloramphenicol and then shifted to the nonpermissive temperature where induction of phi 105 cts23 occurs. dna made after the shift was labeled with a density label, and the distribution of bacterial and phage markers in r ...19734199109
dna repair in bacillus subtilis. ii. activation of the inducible system in competent bacteria.competent b. subtilis are more uv sensitive than the non-competent population of the culture. this increased sensitivity is lose in mutants unable to induce the 'sos system' (reca1,, recg13), in mutants defective in the induction of prophage pbsx (xin), and in late stage competent cells. moreover, bacteriophage phi 105 produced from transfected cells are less restricted on strain yb880 than bacteriophage produced from infected cells. therefore, competent cells (those capable of being transfected ...1977407446
restriction enzyme analysis of bacillus subtilis bacteriophage phi 105 dna.the recognition sites on phi 105 dna for the restriction endonucleases ecori, bg/ii, smai, kpni, ssti, sali, xhoi, ncoi, psti, hindiii, clai, ecorv and mlui have been mapped. the sites for ecori are shown to be different from those published earlier. the dna from phi 105 contains no recognition sites for the endonucleases bamhi and xbai.19846088676
genetic analysis of spo0a and spo0c mutants of bacillus subtilis with a phi 105 prophage merodiploid 8.0-kilobase chromosomal fragment of bacillus subtilis which contained an intact spo0a gene was recloned onto temperate phage phi 105 from the rho 11dspo0a+-1 transducing phage. a specialized transducing phage, phi 105-dspo0a+-1, was constructed and used to transduce the spo0a12 mutant strain 1s9. a spo+ transductant which was a single lysogen of the phi 105dspo0a+-1 transducing phage was isolated. from competent cells of this spo+ transductant was isolated a spo- (spo0a) strain which was imm ...19853926745
molecular cloning and analysis of nucleotide sequence of the bacillus subtilis lysa gene region using b. subtilis phage vectors and a multi-copy plasmid, pub110.a 3.8-kb ecori-fragment containing the lysa gene [diaminopimelate (dap) decarboxylase] of bacillus subtilis has been cloned into b. subtilis phage phi 105 and its nucleotides sequenced. the nucleotide sequence of a 3,762 bp stretch contained three open reading frames (orf1, orf2, and orf3) in one orientation and another open reading frame (orf4) in the opposite orientation. orf2 coded for the lysa gene based on the complementation of a b. subtilis lys auxotroph and on the fact that the predicted ...19911368705
evidence for circular permutation of the prophage genome of bacillus subtilis bacteriophage phi 105.analysis of dna extracted from bacillus subtilis lysogenic for bacteriophage phi 105 was performed by restriction endonuclease digestion and southern hybridization using mature phi 105 dna as a probe. the data revealed that the phi 105 prophage is circularly permuted. digests using the enzymes ecori, smai, psti, and hindiii localized the bacteriophage attachment site (att) to a region 63.4 to 65.7% from the left end of the mature bacteriophage genome. the phi 105 att site-containing smai c, psti ...19863081734
interference of plasmid pcm194 with lysogeny of bacteriophage sp02 in bacillus subtilis.three observations indicated that the 2-megadalton chloramphenicol resistance plasmid pcm194 interferes with sp02 lysogeny of bacillus subtilis. sp02 plaques formed on b. subtilis(pcm194) appeared almost clear, whereas plaques produced on plasmid-free or pub110-containing cells contained large turbid centers. the number of phages spontaneously liberated by b. subtilis(sp02) was increased 10-fold or more when pcm194 was also present in the lysogens. lastly, growth of b. subtilis(sp02, pcm194) for ...19826811554
crosstalk between plasmid vegetative replication and conjugative transfer: repression of the trfa operon by trba of broad host range plasmid rk2.previous deletion and complementation analysis has indicated that the region between trfa and kilbi (trbb) encodes trans-acting factor, designated trba, required for conjugative transfer of broad host range plasmid rk2. in analysing the nucleotide sequence of this region we have discovered a gene encoding a 12 kda polypeptide. the predicted amino acid sequence of this protein shows similarity at its c-terminal to kora from the central control operon of rk2 and at its n-terminal to immunity repre ...19921508679
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