A Simple and Rapid Method for Intra- and Interspecific Transformation of Bacillus subtilis on Solid Media by DNA in Protoplast Lysates

2001 ◽  
Vol 65 (2) ◽  
pp. 446-448 ◽  
Author(s):  
Takashi AKAMATSU ◽  
Hisataka TAGUCHI
Keyword(s):  
Bacillus Subtilis ◽  
Rapid Method ◽  
Solid Media ◽  
Interspecific Transformation

scholarly journals A Rapid Gravimetric Method of Standardising Vaccines

1912 ◽  
Vol 12 (1) ◽  
pp. 49-59
Author(s):  
W. James Wilson ◽  
Charles Dickson
Keyword(s):  
Staphylococcus Aureus ◽  
Rapid Method ◽  
Gravimetric Method ◽  
Platinum Foil ◽  
Solid Media

1. A method is described of rapidly evaporating to dryness bacterial emulsions contained in tarred flasks and from the increase of weight determining the strength of the emulsion.The same apparatus can be used for a variety of purposes, e.g. drying serum, estimating solids in water, milk etc.2. A very rapid method of drying bacteria smeared on a tarred piece of platinum foil, and from the dried residue preparing a vaccine is described.3. A correlation between the number and weight of bacteria in vaccines made from agar cultures of the B. typhosus, B. coli, B. pyocyaneus, Meningococcus (Weichselbaum), Pseudo-Gonococcus, Micrococcus melitensis, B. pneumoniae Friedlander, Staphylococcus aureus and Streptococcus has been established.4. The advantages of the gravimetric method are:(a) That with a sensitive balance constant results are obtained.(b) It is more rapid, and involves no strain ou the eyes.(c) It can be applied to cultures of all bacteria growing on solid media, whether these form uniform emulsions or not, e.g. vaccines of Streptococci, Micrococcus catarrhalis, Diphtheroid organisms, B. tuberculosis, Streptothrices can be accurately standardised. Even small clumps in the vaccine considerably vitiate the results in Wright's method. The chief precaution to take in the gravimetric method is to remove the growth carefully, without breaking the surface of the medium and avoiding the condensation water.(d) It brings into line the dosage of bacterio-proteins with that of other medical remedies.

scholarly journals Retarded swarming motility in Bacillus subtilis NRS-762 and Pseudomonas aeruginosa PRD-10

2018 ◽  
Author(s):  
Wenfa Ng
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Bacillus Subtilis ◽  
Bacterial Species ◽  
Nutritional Stress ◽  
Cellular Level ◽  
Bacterial Cells ◽  
Loss Of Function ◽  
Swarming Motility ◽  
Bacteria Growth ◽  
Solid Media

Coping with nutritional stress is essential for cell survival, of which many strategies at the cellular level lend support for ensuring the survival of the population at a particular habitat. One postulated mechanism is swarming motility in bacterial cells, where, upon depletion of nutrients at a locale, cells would coordinate their movement, synthesize more flagella, and secrete lubricants for moving rapidly across surfaces in search for food. Known to engage in swarming motility, Bacillus subtilis and Pseudomonas aeruginosa are two common bacterial species with versatile metabolism that use the motility mode to colonize new habitats with more favourable environmental and nutritional conditions. However, experimental observations of bacteria growth on a variety of agar media revealed that B. subtilis NRS-762 (ATCC 8473) and P. aeruginosa PRD-10 (ATCC 15442) exhibited retarded swarming motility upon entry into stationary phase on solid media. Specifically, B. subtilis NRS-762 colonies exhibited round, wrinkled morphologies compared to complex filamented swarming patterns common in strains able to engage in swarming motility. On the other hand, P. aeruginosa PRD-10 colonies were round, mucoid, and expanded outwards from the colony centre without extending filaments from the centre; thereby, indicating retarded swarming motility. Thus, impaired cellular machinery for swarming motility or mutated and deleted genes likely account for observed retarded swarming motility in B. subtilis NRS-762 and P. aeruginosa PRD-10. More importantly, observations of small filaments extending radially from an expanded colony of P. aeruginosa PRD-10 grown on minimal salts medium supplemented with yeast extract highlighted possible loss of function of effector molecules that transmit cellular decision at swarming motility into movement, while sensory mechanisms feeding into the motility mechanism remained intact. More broadly, observations of impaired swarming motility in B. subtilis NRS-762 and P. aeruginosa PRD-10 in two species otherwise endowed with the motility mode highlighted that additional triggers for swarming motility are likely present, and the motility mode may have been evolutionary selected for other functions in addition to foraging for food in times of nutritional stress.

scholarly journals Surface-Associated Flagellum Formation and Swarming Differentiation in Bacillus subtilis Are Controlled by the ifm Locus

2004 ◽  
Vol 186 (4) ◽  
pp. 1158-1164 ◽  
Author(s):  
Sonia Senesi ◽  
Emilia Ghelardi ◽  
Francesco Celandroni ◽  
Sara Salvetti ◽  
Eva Parisio ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Solid Medium ◽  
Fold Increase ◽  
Solid Surfaces ◽  
Wild Type ◽  
Liquid Media ◽  
Flagellin Gene ◽  
Swarming Motility ◽  
Solid Media ◽  
Solid Substrates

ABSTRACT Knowledge of the highly regulated processes governing the production of flagella in Bacillus subtilis is the result of several observations obtained from growing this microorganism in liquid cultures. No information is available regarding the regulation of flagellar formation in B. subtilis in response to contact with a solid surface. One of the best-characterized responses of flagellated eubacteria to surfaces is swarming motility, a coordinate cell differentiation process that allows collective movement of bacteria over solid substrates. This study describes the swarming ability of a B. subtilis hypermotile mutant harboring a mutation in the ifm locus that has long been known to affect the degree of flagellation and motility in liquid media. On solid media, the mutant produces elongated and hyperflagellated cells displaying a 10-fold increase in extracellular flagellin. In contrast to the mutant, the parental strain, as well as other laboratory strains carrying a wild-type ifm locus, fails to activate a swarm response. Furthermore, it stops to produce flagella when transferred from liquid to solid medium. Evidence is provided that the absence of flagella is due to the lack of flagellin gene expression. However, restoration of flagellin synthesis in cells overexpressing σD or carrying a deletion of flgM does not recover the ability to assemble flagella. Thus, the ifm gene plays a determinantal role in the ability of B. subtilis to contact with solid surfaces.

scholarly journals Swarming Differentiation and Swimming Motility in Bacillus subtilis Are Controlled by swrA, a Newly Identified Dicistronic Operon

2005 ◽  
Vol 187 (15) ◽  
pp. 5356-5366 ◽  
Author(s):  
Cinzia Calvio ◽  
Francesco Celandroni ◽  
Emilia Ghelardi ◽  
Giuseppe Amati ◽  
Sara Salvetti ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Bacillus Subtilis ◽  
Preliminary Data ◽  
Solid Surfaces ◽  
Cell Contact ◽  
Gene Products ◽  
Swimming Motility ◽  
Solid Media ◽  
Specific Trait

ABSTRACT The number and disposition of flagella harbored by eubacteria are regulated by a specific trait successfully maintained over generations. The genes governing the number of flagella in Bacillus subtilis have never been identified, although the ifm locus has long been recognized to influence the motility phenotype of this microorganism. The characterization of a spontaneous ifm mutant of B. subtilis, displaying diverse degrees of cell flagellation in both liquid and solid media, raised the question of how the ifm locus governs the number and assembly of functional flagella. The major finding of this investigation is the characterization of a newly identified dicistronic operon, named swrA, that controls both swimming motility and swarming differentiation in B. subtilis. Functional analysis of the swrA operon allowed swrAA (previously named swrA [D. B. Kearns, F. Chu, R. Rudner, and R. Losick, Mol. Microbiol. 52:357-369, 2004]) to be the first gene identified in B. subtilis that controls the number of flagella in liquid environments and the assembly of flagella in response to cell contact with solid surfaces. Evidence is given that the second gene of the operon, swrAB, is essential for enabling the surface-adhering cells to undergo swarming differentiation. Preliminary data point to a molecular interaction between the two gene products.

scholarly journals Cosegregation of cell wall and DNA in Bacillus subtilis

1982 ◽  
Vol 152 (3) ◽  
pp. 1231-1240
Author(s):  
J M Schlaeppi ◽  
D Karamata
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Random Distribution ◽  
Chase Period ◽  
Daughter Cells ◽  
Symmetrical Pattern ◽  
Solid Media ◽  
Independent Segregation ◽  
Microscope Autoradiography ◽  
Cell Wall Organization

Cosegregation of cell wall and DNA of a lysis-negative mutant of Bacillus subtilis was examined by continuously labeling (i) cell wall, (ii) DNA, and (iii) both cell wall and DNA. After four to five generations of chase in liquid media it was found by light microscope autoradiography that the numbers of wall segregation units per cell are 29 and 9 in rich and minimal medium, respectively. Under the same conditions the numbers of segregation units of DNA were almost 50% lower: 15 and 5, respectively. Simultaneous labeling of cell wall and DNA (iii) provided figures almost identical to those obtained for cell wall alone, (i), implying cosegregation of the two components. Statistical analysis ruled out their random distribution into daughter cells. Measurements of the positions of grain clusters at the end of the chase period along chains of cells, each derived from a single cell at the beginning of chase, show that cell wall units are localized according to a symmetrical pattern, whereas those of DNA are distributed in an asymmetrical but highly regular way. It appears that of two cell wall units of the same age one only has a strand of DNA attached to it. We present a simple diagrammatic model of cell wall organization and DNA-cell wall association which is compatible with our observations. Finally, we discuss previous experiments pertinent to cosegregation of cell wall and DNA obtained with cells grown on solid media as well as with germinating spores; an explanation for the independent segregation of cell wall and DNA observed in the latter case is advanced.

Rapid method for selecting appropriate solid media for the enumeration of aerobic micro-organisms

1983 ◽  
Vol 54 (3) ◽  
pp. 329-334 ◽  
Author(s):  
J. Richard ◽  
Hanne Møller Andersen ◽  
J.J. Gratadoux
Keyword(s):  
Rapid Method ◽  
Solid Media ◽  
Micro Organisms

scholarly journals A rapid and simple method for Bacillus subtilis transformation on solid media

Microbiology ◽  
1994 ◽  
Vol 140 (7) ◽  
pp. 1613-1617 ◽  
Author(s):  
P. M. HAUSER ◽  
D. KARAMATA
Keyword(s):  
Bacillus Subtilis ◽  
Simple Method ◽  
Solid Media

scholarly journals Genes Involved in Formation of Structured Multicellular Communities by Bacillus subtilis

2004 ◽  
Vol 186 (12) ◽  
pp. 3970-3979 ◽  
Author(s):  
Steven S. Branda ◽  
José Eduardo González-Pastor ◽  
Etienne Dervyn ◽  
S. Dusko Ehrlich ◽  
Richard Losick ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Biofilm Formation ◽  
Regulatory Protein ◽  
Wild Strain ◽  
Laboratory Strain ◽  
Signal Peptidase ◽  
Solid Media ◽  
A Genome ◽  
Wild Strains ◽  
Six Genes

ABSTRACT The spore-forming bacterium Bacillus subtilis is capable of assembling multicellular communities (biofilms) that display a high degree of spatiotemporal organization. Wild strains that have not undergone domestication in the laboratory produce particularly robust biofilms with complex architectural features, such as fruiting-body-like aerial projections whose tips serve as preferential sites for sporulation. To discover genes involved in this multicellular behavior and to do so on a genome-wide basis, we took advantage of a large collection of mutants which have disruptions of most of the uncharacterized genes in the B. subtilis genome. This collection, which was generated with a laboratory strain, was screened for mutants that were impaired in biofilm formation. This subset of mutated genes was then introduced into the wild strain NCIB 3610 to study their effects on biofilm formation in liquid and solid media. In this way we identified six genes that are involved in the development of multicellular communities. These are yhxB (encoding a putative phosphohexomutase that may mediate exopolysaccharide synthesis), sipW (encoding a signal peptidase), ecsB (encoding an ABC transporter subunit), yqeK (encoding a putative phosphatase), ylbF (encoding a regulatory protein), and ymcA (a gene of unknown function). Further analysis revealed that these six genes play different roles in B. subtilis community development.

Sign in / Sign up

Export Citation Format

Share Document