Molecular Cloning in Bacillus subtilis

Gryczan, Thomas J.

doi:10.1016/b978-0-12-222701-1.50015-3

Cited by 14 publications

(7 citation statements)

References 105 publications

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“…Thus biochemical, physiological, morphological and antibiotic resistance tests were included. Although the prevalence of plasmids in bacilli might argue against the use of antibiotic resistance tests, resistance to antibiotics such as chloramphenicol is not commonly plasmid-borne in Bacillus, (Gryczan, 1982). For example, in bacilli chloramphenicol acetyltransferase and streptomycin resistance can be chromosomally encoded (Williams et al, 1981 ;Yousten et al, 1985) and offer useful identification criteria for B. pumilus and B. sphaericus respectively as well as other taxa.…”

Section: Utilization O Fmentioning

confidence: 99%

A Frequency Matrix for Probabilistic Identification of Some Bacilli

Priest

Alexander

1988

Microbiology

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A matrix comprising frequencies for positive results for 44 Bacillus taxa for 30 characters has been constructed. The 44 taxa include most of the common species and several clusters of environmental isolates including those described as B. firmus-B. lentus intermediates. The tests, which were chosen for their high diagnostic value, included some of the traditional tests used for identification of bacilli supplemented with a range of sugar fermentations and other characterization tests. The matrix was evaluated by identifying hypothetical median organisms, cluster representatives and a panel of 23 reference strains. All reference strains achieved Willcox probabilities above 0.995. Fifty-eight environmental isolates were also subjected to the 30 tests and identification was attempted. Forty-one strains (70%) achieved a Willcox probability greater than 0.95, which was considered an acceptable identification, and were assigned to 12 taxa. If the SE of taxonomic distance was also considered in the identification score (an acceptable value being less than 7.0), the number of acceptable identifications was reduced to 34 (59%). It was encouraging that bacteria from garden soils identified to the common species such as B. subtilis, B. cereus and B. licheniformis whereas some of the bacteria from an estuarine habitat were identified as species such as B. firmus which are normally identified with that habitat.

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Section: Utilization O Fmentioning

confidence: 99%

A Frequency Matrix for Probabilistic Identification of Some Bacilli

Priest

Alexander

1988

Microbiology

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“…The complementation observed did not completely restore the germination behavior to that of the wild type. Plasmid pHV33 has been reported to show both segregational and structural instability in E. coli and B. subtilis (7,20). As segregation was prevented by the maintained selection for antibiotic resistance, the heterogeneity of complementation behavior was likely to result from structural instability.…”

Section: Discussionmentioning

confidence: 99%

Identification of three complementation units in the gerA spore germination locus of Bacillus subtilis

Zuberi¹,

Feavers²,

Moir³

1985

J Bacteriol

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The gerA locus, mutations in which affect the germination response of spores to L-alanine and related amino acids, is contained within a 6-kilobase region of DNA cloned in phage and p!asmid vectors. Fragments from this region, subcloned in the shuttle vector pHV33, were introduced into Bacillus subtilis, and their ability to complement chromosomal gerA mutations in a recE4 background was examihed. Although the plasmids were somewhat unstable, it was possible to score complement!tion within spore-containing colonies on nutrient agar by their ability to reduce 2,3,5,-triphenyltetrazolium chloride in an overlay. These studies have assigned the 10 gerA mutations tested to three complementation groups. An analysis of Tn1l000 insertions into the cloned DNA of two relatively stabie plasmids that together encompass the entire gerA region has identified more precisely the location and extent of the complementation units; recombination studies and in vitro mutagenesis'wer,e used to further delineate the extents of two of the units. The evidence suggests that the three complementation units are adjacent and that they are probably capable of separate transcription. * Corresponding author. 756Plasmids. Fragments of B. subtilis DNA were subcloned amphenicol at 3 ,ug ml-' and purified on the nutrient agar described above, again supplemented with chloramphenicol at 3 ,ug ml-'. All cultures were incubated at 37°C, unless otherwise specified.

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“…Unlike the techniques of DNA sequencing and site-directed mutagenesis, the development of recombinant DNA and transposon technologies requires vectors and methods that are frequently useful only among members of one group of bacteria. The reader is encouraged to examine previous reviews of this topic (Lovett, 1981(Lovett, , 1984Gryczan, 1982;. …”

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confidence: 99%

Bacillus

1989

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Molecular Cloning in Bacillus subtilis

Cited by 14 publications

References 105 publications

A Frequency Matrix for Probabilistic Identification of Some Bacilli

A Frequency Matrix for Probabilistic Identification of Some Bacilli

Identification of three complementation units in the gerA spore germination locus of Bacillus subtilis

Bacillus

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