Cloning and Sequencing of a Gene from Bacillus amyloliquefaciens that Complements Mutations of the Sporulation Gene spoIID in Bacillus subtilis

Turner, Sheila M.; Mandelstam, J.

doi:10.1099/00221287-132-11-3025

Cited by 4 publications

(3 citation statements)

References 15 publications

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“…Computer analysis (IDEAS sequence analysis system on the NBRF protein data base, Release 25.0, Kyushu University, Japan) revealed that CwbA exhibits significant amino acid sequence homology with the B. subtilis and B. amyloliquefaciens spoIID gene products (LopezDiaz et al, 1986;Turner & Mandelstam, 1986) (Fig. 3).…”

Section: Resultsmentioning

confidence: 99%

Molecular cloning and sequencing of the upstream region of the major Bacillus subtilis autolysin gene: a modifier protein exhibiting sequence homology to the major autolysin and the spoIID product

Kuroda

Rashid

Sekiguchi

1992

Journal of General Microbiology

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The upstream region of the N-acetylmuramoyl-L-alanine amidase gene (cwfB; a major Baciffus subtifis autolysin) was cloned into Escherichia coli by chromosome walking. Sequencing of the region showed the presence of two open reading frames, one (designated as cwbA) which starts at a UUG codon and encodes a polypeptide of 705 amino acids with an MI of 76725, and the other (designated as Ippx, upstream of cwbA, comprising 102 amino acids and having a signal sequence characteristic of a lipoprotein. Purification of the CwbA protein and determination of its N-terminal amino acid sequence revealed that it contains a presumed signal peptide which is processed after Ala at position 25 from the N-terminal, and that the M, of the mature form is 75000. The amino acid sequences of the N-terminal and C-terminal regions of CwbA were found to be highly homologous with those of the cell wall binding domain of CwlB and the spZZD gene product, respectively. CwbA stimulated the major autolysin activity approximately threefold in nitro. These data indicate that CwbA is the modifier protein of the major autolysin

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Section: Resultsmentioning

confidence: 99%

Molecular cloning and sequencing of the upstream region of the major Bacillus subtilis autolysin gene: a modifier protein exhibiting sequence homology to the major autolysin and the spoIID product

Kuroda

Rashid

Sekiguchi

1992

Journal of General Microbiology

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“…Difficulties have also been encountered when attempting to clone fragments of DNA larger than about 2.5 kbp in certain plasmid vectors (Gryczan & Dubnau, 1982). Plasmids containing inserts often exhibit structural or segregational instability (see, for example, Bron & Luxen, 1985), even in recipients which are Rec- (Tanaka, 1979;Uhlen et al, 1981;Hahn & Dubnau, 1985).…”

Section: Introductionmentioning

confidence: 99%

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

Jones

Errington²

1987

Microbiology

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483A series of improved phage vectors have been constructed, based on Bacillus subtilis bacteriophage 4105, which can be used to clone genes in B. subtilis by direct transfection of protoplasts. The new vectors, designated 4105523, 4105524, 4105527 and 4105528, show frequencies of plaque formation that are equal to those of wild-type 4105. This represents at least a 1 0-fold improvement over 4 105J9, the vector used in previous cloning experiments. Two of the new vectors 4105527 and $105528 incorporate a mutation, cts-52, that renders the prophage temperature inducible. This has made it possible to devise a rapid small-scale procedure for screening progeny phage for the presence of inserted DNA. The usefulness of the new vectors is illustrated in the accompanying paper by cloning more than 20 B. subtilis sporulation genes.

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“…amyloliquefaciens and B. subtilis, finding that they are very similar in both nucleotide sequence and inferred primary protein structure (Turner and Mandelstam, 1986).…”

Section: Sporulationmentioning

confidence: 94%

Microbial Community Structure and Functionality in Ruminants Fed the Probiotic Bacillus amyloliquefaciens H57

Schofield¹

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In the agricultural industry, farmers are continually searching for ways to improve animal production. Developing a more efficient digestive system is one such way that this can be achieved.In domesticated ruminants such as sheep and cattle, the feed material is initially digested in the largest and most prominent compartment of the alimentary canal: the rumen. The rumen is populated by a complex community of microorganisms that are capable of degrading the tough fibrous components of the plant cell wall, releasing fermentable and digestible carbohydrates, proteins, lipids and nucleic acids. Probiotics, defined as "live microorganisms that, when administered in adequate amounts, confer a health benefit on the host", have been used as a means to influence the rumen microbial community to improve the efficiency of feed digestion.In this study we cultivate the probiotic Bacillus amyloliquefaciens strain H57 as an inoculum, to be added to pelleted animal feed in concentrations of 10 9 cfu kg -1 (Chapter 2). To determine the genetic makeup of H57, its genome was sequenced (Chapter 3). The sequence analysis revealed a number of antimicrobial lipopeptides and polyketides, which if expressed in the rumen could significantly alter the rumen microbial community. Other genes of interest within the H57 genome include a number of carbohydrate degrading enzymes as well as those involved with nitrate reduction, which has been recognised as a potential alternative electron acceptor under anaerobic conditions such as the rumen.The H57 inoculum was administered in two animal feeding trials, one using pregnant Dorper ewes and the other using newborn dairy calves. In both instances a significant increase in weight gain was observed in the H57 fed animals compared to the control group. As a means to understand how H57 benefits these animals, the rumen microbial community was profiled by sequencing PCR amplicons of the 16S rRNA gene (Chapter 4). Analysis of the calf rumen microbiome did not reveal any significant differences between the control calves and those that were fed H57 for eight weeks. The only differences in the rumen community occurred between the initial sampling at four weeks and after weaning at eight weeks. In the sheep, however, there was a clear shift in microbial community structure between the control and H57 fed sheep. In both the control and H57 fed sheep the most abundant bacterial populations were Prevotella, although different species were dominant in each treatment group. The H57 fed sheep also had an abundance of Roseburia.Further analysis into the genetic potential and functionality of differentially abundant rumen microorganisms was performed on the sheep samples using metagenomic and metatranscriptomic sequencing. It was found that in the H57 fed sheep the rumen microbial community had an increased expression of hemicellulolytic enzymes, including β-glucosidase/β-galactosidase, α-galactosidase and L-arabinose isomerase. As hemicellulose contributes a significant portion of plant iii cell wall bio...

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Cloning and Sequencing of a Gene from Bacillus amyloliquefaciens that Complements Mutations of the Sporulation Gene spoIID in Bacillus subtilis

Cited by 4 publications

References 15 publications

Molecular cloning and sequencing of the upstream region of the major Bacillus subtilis autolysin gene: a modifier protein exhibiting sequence homology to the major autolysin and the spoIID product

Molecular cloning and sequencing of the upstream region of the major Bacillus subtilis autolysin gene: a modifier protein exhibiting sequence homology to the major autolysin and the spoIID product

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

Microbial Community Structure and Functionality in Ruminants Fed the Probiotic Bacillus amyloliquefaciens H57

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