Construction of Improved Bacteriophage  105 Vectors for Cloning by Transfection in Bacillus subtilis

Jones, D.; Errington, Jeff

doi:10.1099/00221287-133-3-483

Cited by 19 publications

(20 citation statements)

References 36 publications

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“…One CtaA+ lysogen was isolated, purified, and used to prepare phage, designated jl05ctaA'. The ability of P105ctaA' to complement various cta mutations was tested by the cross-streak method described by Jones and Errington (22). Cmr lysogens were scored for their CtaA phenotypes by the TMPD agar plate test.…”

Section: Methodsmentioning

confidence: 99%

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Isolation and sequence of ctaA, a gene required for cytochrome aa3 biosynthesis and sporulation in Bacillus subtilis

Mueller

Taber

1989

J Bacteriol

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Cytochrome aa3 is one of two terminal oxidase complexes in the Bacillus subtilis electron transport chain. A novel genetic strategy was devised which permitted the isolation of B. subtilis mutants lacking cytochrome aa3 by selection for streptomycin-resistant clones which failed to oxidize the artificial electron donor N,N,N',N'-tetramethyl-p-phenylenediamine. Two mutations were studied intensively. Spectroscopic examination showed that each mutant lacked cytochrome aa3; they were also asporogenous

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Section: Methodsmentioning

confidence: 99%

“…Plasmid pAI546 was linearized at its unique BamHI site and cloned into phage vector 1405J23 (22) by transfection of protoplasts of B.…”

Section: Methodsmentioning

confidence: 99%

Isolation and sequence of ctaA, a gene required for cytochrome aa3 biosynthesis and sporulation in Bacillus subtilis

Mueller

Taber

1989

J Bacteriol

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“…All methods for propagation and manipulation of 4105J113 and its derivatives have been described previously (10,12,18). 4105J106, the vector from which 4405J113 was denived, has no BglII restriction sites (10).…”

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The Bacillus subtilis spo0J gene: evidence for involvement in catabolite repression of sporulation

et al. 1991

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Previous observations concerning the ability of the Bacillus subtilis bacteriophages SP10 and PMB12 to suppress mutations in spoOJ and to make wild-type sporulation catabolite resistant suggested that spoOJ had a role in catabolite repression of sporulation. This suggestion was supported in the present report by the ability of the catabolite-resistant sporulation mutation crsF4 to suppress a Tn917 insertion mutation of the B. subtilis spoOJ locus (spoOJ::Tn917fQHU261) in medium without glucose. Although crsF4 and SP10 made wild-type B. subtilis sporulation catabolite resistant, neither crsF4 nor SP10 caused a mutant with spoOJ::Tn917Q1HU261 to sporulate in medium with glucose. Sequencing the spoOJ locus revealed an open reading frame that was 179 codons in length. Disruption of the open reading frame resulted in a sporulation-negative (Spo-) phenotype that was similar to those of other spoOJ mutations. Analysis of the deduced amino acid sequence of the spoOJ locus indicated that the spoOJ gene product contains an ot-helix-turn-a-helix unit similar to the motif found in Cro-like DNA-binding proteins.Bacillus subtilis sporulation is a model system that is used to study procaryotic gene expression and cellular differentiation as responses to environmental stimuli. Sporulation is subject to catabolite repression; i.e., the presence of glucose or other readily metabolized carbon sources inhibits sporulation by wild-type cells (33). Initiation of sporulation is controlled by at least seven genes, spoOA, spoOB, spoOE, spoOF, spoOH, spoOJ, and spoOK (23). Glucose represses transcription of spoOA and spoOF (3, 43). However, it is not known how availability of nutrients regulates initiation of sporulation. Several spoO genes have been sequenced (5,9,14,15,30,41). It is evident from this work that the spoOH gene codes for a sigma subunit of RNA polymerase and that some spoO genes are responsible for sensing environmental conditions. The spoOA and spoOF gene products are homologous to the effector molecules of the two-component response regulator systems that have been described for a variety of bacteria (15,41 PMB12 and SP10 are also able to suppress the Spo-and oligosporogenic phenotypes of various spoOJ mutations (6,20,34). As a result, both PMB12-infected and SP10-infected spoOJ mutants sporulate at a significantly higher frequency than uninfected spoOJ mutants. The observations that PMB12-and SP10-infected bacteria display catabolite-resistant sporulation and that these bacteriophages suppress spoOJ mutations suggest that spoOJ has a role in catabolite repression of sporulation.The spoOJ locus was originally thought to be represented by two mutations, spoOJ87 and spoOJ93 (17). However, spoIIA and spaIID are expressed in a strain with spoOJ87, whereas spoOJ93 blocks expression of these genes (7, 13). The wild-type alleles of both mutations have been cloned into bacteriophage 0105 vectors (10,12 spoOJ93 (45).The data presented in this report provide additional evidence for the involvement of spoOJ in catabolite repressi...

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“…This is consistent with the spoIIIC transcription unit beginning between the leftmost RsaI site and PvuI site and ending before the AccI site. As a further check on the location of the spoIIIC' gene, the 1.3-kbp HindIlI fragment of pSGMU54 was subcloned into the phage vector 4105J23 (19 …”

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confidence: 99%

“…Plasmid pSGMU54 was linearized at its unique BamHI site and cloned into phage vector 41i05J23 (19) by transfection of protoplasts of B. subtilis 168 (7,19) followed by selection for transduction of the same strain to resistance to chloramphenicol (5 ,ug/ml). RESULTS spoIIIC94 mutation is a large deletion.…”

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confidence: 99%

Transcriptional regulation and structure of the Bacillus subtilis sporulation locus spoIIIC

et al. 1988

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. 146: [430][431][432] 1981) by using as an assay transformation of the mutant allele spoIIIC94 to the wild type. Regulation of the spolliC locus was studied by hybridization of cloned spolliC DNA to RNA pulse-labeled at various times during growth and sporulation. The relative rate of transcription of the spolIlC locus was highest 3 h after the end of growth. The DNA sequence of the spoIlIC transcription unit indicated the coding capacity for a small protein (138 amino acids) having significant similarity with one domain of RNA polymerase sigma factors. Interruption of this coding sequence by an insertion mutation caused cells to become Spo-.The gram-positive soil bacterium Bacillus subtilis responds to nutrient deprivation by undergoing a series of metabolic and morphological changes that culminate in formation of a dormant endospore. These changes follow a temporal pattern that has been well defined. More than 50 genetic loci have been identified at which mutations (called spo mutations) block sporulation without inhibiting growth (22). Although the specific functions of these sporulation genes are generally unknown, the mutations cause blockage at identifiable morphological stages. Several of these sporulation genes have been isolated recently (1,7,8,10,16,28,35,37). In some cases it has been possible to demonstrate specific transcription of particular sporulation genes at different times during sporulation (28, 32).The study of other genes, which were isolated on the basis of their specific expression during sporulation, has provided strong evidence supporting the hypothesis (20,21) that sets of genes are switched on or off at particular times during sporulation by sequential replacement of the sigma factor of RNA polymerase. This component of RNA polymerase is known to play a critical role in determining promoter specificity. Through the use of such cloned genes four minor vegetative forms of RNA polymerase, Ea28, Ea30, Eu32, and Eu37, and one sporulation-specific form, Eur29, have been identified (5,6,12,14,15,18). These studies demonstrated the specific transcriptional activities of these forms of RNA polymerase by using various genes as templates for in vitro experiments. These templates have not generally been genes whose products are clearly required for sporulation.To test the generality of the sigma-replacement hypothesis for regulation of spo gene expression, we have been isolating additional genes whose products are required for sporulation. We previously reported the isolation and transcriptional analysis of the spoIID gene (28), and we report here initial studies on the cloned spoIIIC locus.

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Construction of Improved Bacteriophage 105 Vectors for Cloning by Transfection in Bacillus subtilis

Cited by 19 publications

References 36 publications

Isolation and sequence of ctaA, a gene required for cytochrome aa3 biosynthesis and sporulation in Bacillus subtilis

Isolation and sequence of ctaA, a gene required for cytochrome aa3 biosynthesis and sporulation in Bacillus subtilis

The Bacillus subtilis spo0J gene: evidence for involvement in catabolite repression of sporulation

Transcriptional regulation and structure of the Bacillus subtilis sporulation locus spoIIIC

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