Mini-Tn7 Insertion in an Artificial attTn7 Site Enables Depletion of the Essential Master Regulator CtrA in the Phytopathogen Agrobacterium tumefaciens.


UNLABELLED:Mechanistic studies of many processes in Agrobacterium tumefaciens have been hampered by a lack of genetic tools for characterization of essential genes. In this study, we used a Tn7-based method for inducible control of transcription from an engineered site on the chromosome. We demonstrate that this method enables tighter control of inducible promoters than plasmid-based systems and can be used for depletion studies. The method enables the construction of depletion strains to characterize the roles of essential genes in A. tumefaciens Here, we used the strategy to deplete the alphaproteobacterial master regulator CtrA and found that depletion of this essential gene results in dramatic rounding of cells, which become nonviable. IMPORTANCE:Agrobacterium tumefaciens is a bacterial plant pathogen and natural genetic engineer. Thus, studies of essential processes, including cell cycle progression, DNA replication and segregation, cell growth, and division, may provide insights for limiting disease or improving biotechnology applications.


Appl Environ Microbiol


Figueroa-Cuilan W,Daniel JJ,Howell M,Sulaiman A,Brown PJ




Has Abstract


2016-07-29 00:00:00














  • Intracellular accumulation of polyphosphate by the yeast Candida humicola G-1 in response to acid pH.

    abstract::Cells of a newly isolated environmental strain of Candida humicola accumulated 10-fold more polyphosphate (polyP), during active growth, when grown in complete glucose-mineral salts medium at pH 5.5 than when grown at pH 7.5. Neither phosphate starvation, nutrient limitation, nor anaerobiosis was required to induce po...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: McGrath JW,Quinn JP

    更新日期:2000-09-01 00:00:00

  • Constraints on anaerobic respiration in the hyperthermophilic Archaea Pyrobaculum islandicum and Pyrobaculum aerophilum.

    abstract::Pyrobaculum islandicum uses iron, thiosulfate, and elemental sulfur for anaerobic respiration, while Pyrobaculum aerophilum uses iron and nitrate; however, the constraints on these processes and their physiological mechanisms for iron and sulfur reduction are not well understood. Growth rates on sulfur compounds are h...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Feinberg LF,Srikanth R,Vachet RW,Holden JF

    更新日期:2008-01-01 00:00:00

  • Water disinfection with the hydrogen peroxide-ascorbic acid-copper (II) system.

    abstract::Treatment of secondary effluents with hydrogen peroxide (10 mg/liter)-ascorbic acid (10 mg/liter)-Cu2+ (0.5 mg/liter) for 60 min resulted in around 99% reduction of the initial plate count. Hydrogen peroxide could be replaced by other peroxygen compounds; ascorbic acid could be replaced by other reducing agents, of wh...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Ragab-Depre NJ

    更新日期:1982-09-01 00:00:00

  • Anaerobic incubation of membrane filter cultures for improved detection of fecal coliforms from recreational waters.

    abstract::Anaerobic incubation of membrane filter cultures significantly enhanced detection of fecal coliforms in surface-water samples from recreational beaches. In contrast to standard aerobic incubation, anaerobic incubation suppressed overgrowth of masking, noncoliform bacteria but did not increase the frequency of fecal co...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Doyle JD,Tunnicliff B,Brickler SK,Kramer RE,Sinclair NA

    更新日期:1984-08-01 00:00:00

  • Bacterial leaching of metal sulfides proceeds by two indirect mechanisms via thiosulfate or via polysulfides and sulfur

    abstract::The acid-insoluble metal sulfides FeS2, MoS2, and WS2 are chemically attacked by iron(III) hexahydrate ions, generating thiosulfate, which is oxidized to sulfuric acid. Other metal sulfides are attacked by iron(III) ions and by protons, resulting in the formation of elemental sulfur via intermediary polysulfides. Sulf...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Schippers A,Sand W

    更新日期:1999-01-01 00:00:00

  • Evaluation of MK filters for recovery of enteroviruses from tap water.

    abstract::The MK filter is an electropositively charged filter that can be used to concentrate enteroviruses from large volumes (400 to 1,000 liters) of water. This filter is less expensive than the commonly used 1MDS electropositive filter. In this study, we compared the recovery of poliovirus 1 (PV1) and that of coxsackieviru...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Ma JF,Naranjo J,Gerba CP

    更新日期:1994-06-01 00:00:00

  • Chloromethane utilization gene cluster from Hyphomicrobium chloromethanicum strain CM2(T) and development of functional gene probes to detect halomethane-degrading bacteria.

    abstract::Hyphomicrobium chloromethanicum CM2(T), an aerobic methylotrophic member of the alpha subclass of the class proteobacteria, can grow with chloromethane as the sole carbon and energy source. H. chloromethanicum possesses an inducible enzyme system for utilization of chloromethane, in which two polypeptides (67-kDa CmuA...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: McAnulla C,Woodall CA,McDonald IR,Studer A,Vuilleumier S,Leisinger T,Murrell JC

    更新日期:2001-01-01 00:00:00

  • Lysogenic strains of lactic Acid streptococci and lytic spectra of their temperate bacteriophages.

    abstract::A total of 113 strains of mesophilic strains lactic streptococci of the species Streptococcus lactis, S. lactis subsp. diacetilactis, and S. cremoris, chosen from 291 strains that had been previously classified into six groups on the basis of their sensitivity to 132 virulent phages, were subjected to induction with m...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Reyrolle J,Chopin MC,Letellier F,Novel G

    更新日期:1982-02-01 00:00:00

  • Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans.

    abstract::Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Cohen-Rosenzweig C,Guan Z,Shaanan B,Eichler J

    更新日期:2014-01-01 00:00:00

  • Rumen methanogenic genotypes differ in abundance according to host residual feed intake phenotype and diet type.

    abstract::Methane is an undesirable end product of rumen fermentative activity because of associated environmental impacts and reduced host feed efficiency. Our study characterized the rumen microbial methanogenic community in beef cattle divergently selected for phenotypic residual feed intake (RFI) while offered a high-forage...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Carberry CA,Waters SM,Kenny DA,Creevey CJ

    更新日期:2014-01-01 00:00:00

  • Characteristics of beta-lactamase-inhibiting proteins from Streptomyces exfoliatus SMF19.

    abstract::Streptomyces exfoliatus SMF19 produced two types of extracellular beta-lactamase-inhibiting proteins, 48 (beta LIP-I) and 33 (beta LIP-II) kDa. The inhibition mode of beta LIP-I on Bacto Penase and benzylpenicillin as substrates was uncompetitive (i.e., both Km and Vmax were changed), while that of beta LIP-II was non...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Kim MK,Lee KJ

    更新日期:1994-03-01 00:00:00

  • Detection of Vibrio vulnificus biotypes 1 and 2 in eels and oysters by PCR amplification.

    abstract::DNA extraction procedures and PCR conditions to detect Vibrio vulnificus cells naturally occurring in oysters were developed. In addition, PCR amplification of V. vulnificus from oysters seeded with biotype 1 cells was demonstrated. By the methods described, V. vulnificus cells on a medium (colistin-polymyxin B-cellob...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Coleman SS,Melanson DM,Biosca EG,Oliver JD

    更新日期:1996-04-01 00:00:00

  • Wood-destroying soft rot fungi in the historic expedition huts of Antarctica.

    abstract::Three expedition huts in the Ross Sea region of Antarctica, built between 1901 and 1911 by Robert F. Scott and Ernest Shackleton, sheltered and stored the supplies for up to 48 men for 3 years during their explorations and scientific investigation in the South Pole region. The huts, built with wood taken to Antarctica...

    journal_title:Applied and environmental microbiology

    pub_type: 历史文章,杂志文章


    authors: Blanchette RA,Held BW,Jurgens JA,McNew DL,Harrington TC,Duncan SM,Farrell RL

    更新日期:2004-03-01 00:00:00

  • Change in Size of Chromatium minus Cells in Relation to Growth Rate, Sulfur Content, and Photosynthetic Activity: A Comparison of Pure Cultures and Field Populations.

    abstract::The size frequency distribution of planktonic cells of purple sulfur phototrophic bacteria was measured at several depths in a bacterial layer of Lake Cisó (Spain). The bacterioplankton was dominated by Chromatium minus (87 to 94% of the total biomass). The largest cells of C. minus were found in the top part of the b...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Montesinos E

    更新日期:1987-04-01 00:00:00

  • Life at the energetic edge: kinetics of circumneutral iron oxidation by lithotrophic iron-oxidizing bacteria isolated from the wetland-plant rhizosphere.

    abstract::Batch cultures of a lithotrophic Fe(II)-oxidizing bacterium, strain BrT, isolated from the rhizosphere of a wetland plant, were grown in bioreactors and used to determine the significance of microbial Fe(II) oxidation at circumneutral pH and to identify abiotic variables that affect the partitioning between microbial ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Neubauer SC,Emerson D,Megonigal JP

    更新日期:2002-08-01 00:00:00

  • A simple filtration method to remove plankton-associated Vibrio cholerae in raw water supplies in developing countries.

    abstract::Plankton to which cells of Vibrio cholerae O1 and/or O139 were attached was introduced into 0.5% Instant Ocean microcosms maintained at 25 degrees C. The bulk of the plankton and associated particulates was removed with a filter constructed from either nylon net and one of several different types of sari material, the...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Huq A,Xu B,Chowdhury MA,Islam MS,Montilla R,Colwell RR

    更新日期:1996-07-01 00:00:00

  • Data Acceptance Criteria for Standardized Human-Associated Fecal Source Identification Quantitative Real-Time PCR Methods.

    abstract::There is growing interest in the application of human-associated fecal source identification quantitative real-time PCR (qPCR) technologies for water quality management. The transition from a research tool to a standardized protocol requires a high degree of confidence in data quality across laboratories. Data quality...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Shanks OC,Kelty CA,Oshiro R,Haugland RA,Madi T,Brooks L,Field KG,Sivaganesan M

    更新日期:2016-04-18 00:00:00

  • Effect of culture conditions on microorganism identification by matrix-assisted laser desorption ionization mass spectrometry.

    abstract::Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) has been used to identify bacteria based upon protein signatures. This research shows that while some different proteins are produced by vegetative bacteria when they are cultured in different growth media, positive identificat...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Valentine N,Wunschel S,Wunschel D,Petersen C,Wahl K

    更新日期:2005-01-01 00:00:00

  • Isolation of Azospirillum lipoferum 4T Tn5 Mutants Affected in Melanization and Laccase Activity.

    abstract::Azospirillum lipoferum 4T has original properties such as nonmotility, melanin synthesis, and laccase activity. Following random Tn5 mutagenesis in A. lipoferum 4T, we obtained 10 mutants which were affected in melanization and laccase activity. The class 1 mutants, with intermediate levels of laccase activity, showed...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Faure D,Bouillant ML,Bally R

    更新日期:1994-09-01 00:00:00

  • Characteristics of a Streptomyces coelicolor A3(2) extracellular protein targeting chitin and chitosan.

    abstract::Upstream of the Streptomyces coelicolor A3(2) chitinase G gene, a small gene (named chb3) is located whose deduced product shares 37% identical amino acids with the previously described CHB1 protein from Streptomyces olivaceoviridis. The chb3 gene and its upstream region were cloned in a multicopy vector and transform...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Saito A,Miyashita K,Biukovic G,Schrempf H

    更新日期:2001-03-01 00:00:00

  • NAH plasmid-mediated catabolism of anthracene and phenanthrene to naphthoic acids.

    abstract::Pseudomonas fluorescens 5R contains an NAH7-like plasmid (pKA1), and P. fluorescens 5R mutant 5RL contains a bioluminescent reporter plasmid (pUTK21) which was constructed by transposon mutagenesis. Polymerase chain reaction mapping confirmed the localization of lux transposon Tn4431 300 bp downstream from the start o...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Menn FM,Applegate BM,Sayler GS

    更新日期:1993-06-01 00:00:00

  • A Synechococcus PglnA::luxAB fusion for estimation of nitrogen bioavailability to freshwater cyanobacteria.

    abstract::In contrast to extensive studies of phosphorus, widely considered the main nutrient limiting phytoplankton biomass in freshwater ecosystems, there have been few studies on the role of nitrogen in controlling phytoplankton populations. This situation may be due partly to the complexity in estimating its utilization and...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Gillor O,Harush A,Hadas O,Post AF,Belkin S

    更新日期:2003-03-01 00:00:00

  • Interaction of Aeromonas strains with lactic acid bacteria via Caco-2 cells.

    abstract::The genus Aeromonas includes some species that have now been identified as human pathogens of significant medical importance. We investigated the ability of 13 selected Aeromonas strains belonging to nine species isolated from clinical cases (n = 5), environmental waters (n = 5), and fish (n = 3) to adhere to and tran...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Hatje E,Neuman C,Katouli M

    更新日期:2014-01-01 00:00:00

  • Engineering a Saccharomyces cerevisiae wine yeast that exhibits reduced ethanol production during fermentation under controlled microoxygenation conditions.

    abstract::We recently showed that expressing an H(2)O-NADH oxidase in Saccharomyces cerevisiae drastically reduces the intracellular NADH concentration and substantially alters the distribution of metabolic fluxes in the cell. Although the engineered strain produces a reduced amount of ethanol, a high level of acetaldehyde accu...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Heux S,Sablayrolles JM,Cachon R,Dequin S

    更新日期:2006-09-01 00:00:00

  • Fluorescence-activated cell sorting of specific affibody-displaying staphylococci.

    abstract::Efficient enrichment of staphylococcal cells displaying specific heterologous affinity ligands on their cell surfaces was demonstrated by using fluorescence-activated cell sorting. Using bacterial surface display of peptide or protein libraries for the purpose of combinatorial protein engineering has previously been i...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Wernérus H,Samuelson P,Ståhl S

    更新日期:2003-09-01 00:00:00

  • Characterization of Pseudomonas aeruginosa growth on O-acylcarnitines and identification of a short-chain acylcarnitine hydrolase.

    abstract::To survive in various environments, from host tissue to soil, opportunistic bacterial pathogens must be metabolically flexible and able to use a variety of nutrient sources. We are interested in Pseudomonas aeruginosa's catabolism of quaternary amine compounds that are prevalent in association with eukaryotes. Carniti...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Meadows JA,Wargo MJ

    更新日期:2013-06-01 00:00:00

  • Listeria phage and phage tail induction triggered by components of bacterial growth media (phosphate, LiCl, nalidixic acid, and acriflavine).

    abstract::The detection of Listeria monocytogenes from food is currently carried out using a double enrichment. For the ISO methodology, this double enrichment is performed using half-Fraser and Fraser broths, in which the overgrowth of L. innocua can occur in samples where both species are present. In this study, we analyzed t...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Lemaître JP,Duroux A,Pimpie R,Duez JM,Milat ML

    更新日期:2015-03-01 00:00:00

  • Characterization of lipopolysaccharides present in settled house dust.

    abstract::The 3-hydroxy fatty acids (3-OHFAs) in lipopolysaccharides (LPS) play an important role in determining endotoxin activity, and childhood exposure to endotoxin has recently been associated with reduced risk of atopic diseases. To characterize the 3-OHFAs in house dust (HD), we used gas chromatography-mass spectrometry ...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Park JH,Szponar B,Larsson L,Gold DR,Milton DK

    更新日期:2004-01-01 00:00:00

  • Generation of a superoxide dismutase (SOD)-deficient mutant of Campylobacter coli: evidence for the significance of SOD in Campylobacter survival and colonization.

    abstract::The microaerophilic nature of Campylobacter species implies an inherent sensitivity towards oxygen and its reduction products, particularly the superoxide anion. The deleterious effects of exposure to superoxide radicals are counteracted by the activity of superoxide dismutase (SOD). We have shown previously that Camp...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Purdy D,Cawthraw S,Dickinson JH,Newell DG,Park SF

    更新日期:1999-06-01 00:00:00

  • Effect of lactose monolaurate on pathogenic and nonpathogenic bacteria.

    abstract::The antimicrobial activities of sucrose monolaurate and a novel ester, lactose monolaurate (LML), were tested. Gram-positive bacteria were more susceptible than Gram-negative bacteria to both esters. The minimal bactericidal concentrations of LML were 5 to 9.5 mM for Listeria monocytogenes isolates and 0.2 to 2 mM for...

    journal_title:Applied and environmental microbiology

    pub_type: 杂志文章


    authors: Wagh A,Shen S,Shen FA,Miller CD,Walsh MK

    更新日期:2012-05-01 00:00:00