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Venkova, Tatiana, Juárez, Antonio, Espinosa, Manuel, (2017). Editorial: Modulating prokaryotic lifestyle by DNA-binding proteins: Learning from (apparently) simple systems Frontiers in Molecular Biosciences 3, Article 86

Within the research in Molecular Biology, one important field along the years has been the analyses on how prokaryotes regulate the expression of their genes and what the consequences of these activities are. Prokaryotes have attracted the interests of researchers not only because the processes taking place in their world are important to cells, but also because many of the effects often can be readily measured, both at the single cell level and in large populations. Contributing to the interest of the present topic is the fact that modulation of gene activity involves the sensing of intra- and inter-cellular conditions, DNA binding and DNA dynamics, and interaction with the replication/transcription machinery of the cell. All of these processes are fundamental to the operation of a biological entity and they condition its lifestyle. Further, the discoveries achieved in the bacterial world have been of ample use in eukaryotes. In addition to the fundamental interest of understanding modulation of prokaryotic lifestyle by DNA-binding proteins, there is an added interest from the healthcare point of view. As it is well-known the antibiotic-resistance strains of pathogenic bacteria are a major world problem, so that there is an urgent need of innovative approaches to tackle it. Human and animal infectious diseases impose staggering costs worldwide in terms of loss of human life and livestock, diminished productivity, and the heavy economic burden of disease. The global dimension of international trade, personal travel, and population migration expands at an ever-accelerating rate. This increasing mobility results in broader and quicker dissemination of bacterial pathogens and in rapid spread of antibiotic resistance. The majority of the newly acquired resistances are horizontally spread among bacteria of the same or different species by processes of lateral (horizontal) gene transfer, so that discovery of new antibiotics is not the definitive solution to fighting infectious diseases. There is an absolute need of finding novel alternatives to the “classical” approach to treat infections by bacterial pathogens, and these new ways must include the exploration and introduction of novel antibacterials, the development of alternative strategies, and the finding of novel bacterial targets. However, all these approaches will result in a stalemate if we, researchers, are not able to achieve a better understanding of the mechanistic processes underlying bacterial gene expression. It is, then, imperative to continue gaining insight into the basic mechanisms by which bacterial cells regulate the expression of their genes. That is why our Research Topic hosted by Frontiers in Molecular Biosciences was timely, and the output of it offers novel and up-to-date points of view to the “simple” bacterial world.

Keywords: DNA-protein interactions, Gene regulation in Prokaryotes, Replication control, Regulation of Bacterial Gene Expression, Global Regulatory Networks


Stanton, Morgan M., Sánchez, Samuel, (2017). Pushing bacterial biohybrids to In Vivo Applications Trends in Biotechnology In Press Corrected Proof

Bacterial biohybrids use the energy of bacteria to manipulate synthetic materials with the goal of solving biomedical problems at the micro- and nanoscale. We explore current in vitro studies of bacterial biohybrids, the first attempts at in vivo biohybrid research, and problems to be addressed for the future.

Keywords: Bacteria, Biohybrid, Microswimmers, Micromotors, Drug delivery


Matalonga, J., Glaria, E., Bresque, M., Escande, C., Carbó, J. M., Kiefer, K., Vicente, R., León, T. E., Beceiro, S., Pascual-García, M., Serret, J., Sanjurjo, L., Morón-Ros, S., Riera, A., Paytubi, S., Juarez, A., Sotillo, F., Lindbom, L., Caelles, C., Sarrias, M. R., Sancho, J., Castrillo, A., Chini, E. N., Valledor, A. F., (2017). The nuclear receptor LXR limits bacterial infection of host macrophages through a mechanism that impacts cellular NAD metabolism Cell Reports 18, (5), 1241-1255

Macrophages exert potent effector functions against invading microorganisms but constitute, paradoxically, a preferential niche for many bacterial strains to replicate. Using a model of infection by Salmonella Typhimurium, we have identified a molecular mechanism regulated by the nuclear receptor LXR that limits infection of host macrophages through transcriptional activation of the multifunctional enzyme CD38. LXR agonists reduced the intracellular levels of NAD+ in a CD38-dependent manner, counteracting pathogen-induced changes in macrophage morphology and the distribution of the F-actin cytoskeleton and reducing the capability of non-opsonized Salmonella to infect macrophages. Remarkably, pharmacological treatment with an LXR agonist ameliorated clinical signs associated with Salmonella infection in vivo, and these effects were dependent on CD38 expression in bone-marrow-derived cells. Altogether, this work reveals an unappreciated role for CD38 in bacterial-host cell interaction that can be pharmacologically exploited by activation of the LXR pathway.

Keywords: Bacterial infection, CD38, Cytoskeleton, LXR, Macrophage, NAD, Nuclear receptor


Simmchen, Juliane, Baeza, Alejandro, Miguel-Lopez, Albert, Stanton, Morgan M., Vallet-Regi, Maria, Ruiz-Molina, Daniel, Sánchez, Samuel, (2017). Dynamics of novel photoactive AgCl microstars and their environmental applications ChemNanoMat 3, (1), 65-71

In the field of micromotors many efforts are taken to find a substitute for peroxide as fuel. While most approaches turn towards other toxic high energy chemicals such as hydrazine, we introduce an energy source that is widely used in nature: light. Light is an ideal source of energy and some materials, such as AgCl, have the inherent property to transform light energy for chemical processes, which can be used to achieve propulsion. In the case of silver chloride, one observed process after light exposure is surface modification which leads to the release of ions generating chemo-osmotic gradients. Here we present endeavours to use those processes to propel uniquely shaped micro objects of micro star morphology with a high surface to volume ratio, study their dynamics and present approaches to go towards real environmental applications.

Keywords: Self-propellers, Silver chloride, Environmental applications, Photoactive colloids, Anti bacterial


Schwab, S., Lehmann, J., Lutz, P., Jansen, C., Appenrodt, B., Lammert, F., Strassburg, C. P., Spengler, U., Nischalke, H. D., Trebicka, J., (2017). Influence of genetic variations in the SOD1 gene on the development of ascites and spontaneous bacterial peritonitis in decompensated liver cirrhosis European Journal of Gastroenterology and Hepatology 29, (7), 800-804

Background The balance between generation and elimination of reactive oxygen species by superoxide dismutase (SOD) is crucially involved in the pathophysiology of liver cirrhosis. Reactive oxygen species damage cells and induce inflammation/fibrosis, but also play a critical role in immune defense from pathogens. As both processes are involved in the development of liver cirrhosis and its complications, genetic variation of the SOD1 gene was investigated. Patients and methods Two SOD1 single nucleotide polymorphisms (rs1041740 and rs3844942) were analyzed in 49 cirrhotic patients undergoing liver transplantation. In addition, 344 cirrhotic patients with ascites were analyzed in a cohort of 521 individuals in terms of the relationship of these polymorphisms with spontaneous bacterial peritonitis (SBP). Results Although rs3844942 showed no associations with complications of cirrhosis, we observed a significant association between rs1041740 and the presence of ascites and SBP in the discovery cohort of patients with cirrhosis. Importantly, the association with SBP was not confirmed in the validation cohort of patients with ascites. By contrast, a trend toward lower SBP rates was observed in carriers of rs1041740. In this cohort, rs1041740 was not associated with survival. Conclusion These data suggest a complex role of SOD1 in different processes leading to complications of liver cirrhosis. rs1041740 might be associated with the development of ascites and possibly plays a role in SBP once ascites has developed.

Keywords: Ascites, Genetic polymorphism, Liver cirrhosis, Reactive oxygen stress, Spontaneous bacterial peritonitis, Superoxide dismutases


Noguera-Ortega, Estela, Secanella-Fandos, Silvia, Eraña, Hasier, Gasión, Jofre, Rabanal, Rosa M., Luquin, Marina, Torrents, Eduard, Julián, Esther, (2016). Nonpathogenic Mycobacterium brumae inhibits bladder cancer growth in vitro, ex vivo, and in vivo European Urology Focus 2, (1), 67-76

Background Bacillus Calmette-Guérin (BCG) prevents tumour recurrence and progression in non–muscle-invasive bladder cancer (BC). However, common adverse events occur, including BCG infections. Objective To find a mycobacterium with similar or superior antitumour activity to BCG but with greater safety. Design In vitro, ex vivo, and in vivo comparisons of the antitumour efficacy of nonpathogenic mycobacteria and BCG. Intervention The in vitro antitumour activity of a broad set of mycobacteria was studied in seven different BC cell lines. The most efficacious was selected and its ex vivo capacity to activate immune cells and its in vivo antitumour activity in an orthotopic murine model of BC were investigated. Outcome measurements and statistical analysis Growth inhibition of BC cells was the primary outcome measurement. Parametric and nonparametric tests were use to analyse the in vitro results, and a Kaplan-Meier test was applied to measure survival in mycobacteria-treated tumour-bearing mice. Results and limitations Mycobacterium brumae is superior to BCG in inhibiting low-grade BC cell growth, and has similar effects to BCG against high-grade cells. M. brumae triggers an indirect antitumour response by activating macrophages and the cytotoxic activity of peripheral blood cells against BC cells. Although no significant differences were observed between BCG and M. brumae treatments in mice, M. brumae treatment prolonged survival in comparison to BCG treatment in tumour-bearing mice. In contrast to BCG, M. brumae does not persist intracellularly or in tumour-bearing mice, so the risk of infection is lower. Conclusions Our preclinical data suggest that M. brumae represents a safe and efficacious candidate as a therapeutic agent for non–muscle-invasive BC. Patient summary We investigated the antitumour activity of nonpathogenic mycobacteria in in vitro and in vivo models of non–muscle-invasive bladder cancer. We found that Mycobacterium brumae effectively inhibits bladder cancer growth and helps the host immune system to eradicate cancer cells, and is a promising agent for antitumour immunotherapy.

Keywords: Animal models, Bacillus Calmette-Guérin, Cytokines, Immunomodulation, Immunotherapy, Mycobacteria, Urothelial cell line


Stanton, Morgan M., Simmchen, Juliane, Ma, Xing, Miguel-López, Albert, Sánchez, Samuel, (2016). Biohybrid Janus motors driven by Escherichia coli Advanced Materials Interfaces 3, (2), 1500505

There has been a significant interest in the development of microswimmers for medical drug and cargo delivery, but the majority of current micromotors rely on toxic fuel sources and materials in their design making them irrelevant for biomedical applications. Bacteria represent an excellent motor alternative, as they are powered using their surrounding biological fluids. For a motile, biohybrid swimmer, Escherichia coli (E. coli) are integrated onto metal capped, polystyrene (PS) Janus particles. Fabrication of the biohybrid is rapid and simple for a microswimmer capable of magnetic guidance and ferrying an anticancer agent. Cell adhesion is regulated as E. coli adheres only to the particle's metal caps allowing the PS surface to be utilized for drug attachment, creating a multifunctional system. E. coli adhesion is investigated on multiple metal caps (Pt, Fe, Ti, or Au) and displays a strong preference to attach to Pt surfaces over other metals. Surface hydrophobicity and surface charge are examined to interpret the cell specific adhesion on the Janus particles. The dual capability of the biohybrid to have guided cell adhesion and localized drug attachment allows the swimmer to have multiple applications for biomedical microswimmers, future bacteria-interface systems, and micro-biorobots.

Keywords: Bacteria adhesion, Biohybrids, Escherichia coli, Janus particles, Microswimmers


Páez-Avilés, C., Juanola-Feliu, E., Punter-Villagrasa, J., Del Moral Zamora, B., Homs-Corbera, A., Colomer-Farrarons, J., Miribel-Català , P. L., Samitier, J., (2016). Combined dielectrophoresis and impedance systems for bacteria analysis in microfluidic on-chip platforms Sensors 16, (9), 1514

Bacteria concentration and detection is time-consuming in regular microbiology procedures aimed to facilitate the detection and analysis of these cells at very low concentrations. Traditional methods are effective but often require several days to complete. This scenario results in low bioanalytical and diagnostic methodologies with associated increased costs and complexity. In recent years, the exploitation of the intrinsic electrical properties of cells has emerged as an appealing alternative approach for concentrating and detecting bacteria. The combination of dielectrophoresis (DEP) and impedance analysis (IA) in microfluidic on-chip platforms could be key to develop rapid, accurate, portable, simple-to-use and cost-effective microfluidic devices with a promising impact in medicine, public health, agricultural, food control and environmental areas. The present document reviews recent DEP and IA combined approaches and the latest relevant improvements focusing on bacteria concentration and detection, including selectivity, sensitivity, detection time, and conductivity variation enhancements. Furthermore, this review analyses future trends and challenges which need to be addressed in order to successfully commercialize these platforms resulting in an adequate social return of public-funded investments.

Keywords: Bacteria, Dielectrophoresis, Impedance, Microfluidics, On-chip


Barniol-Xicota, M., Escandell, A., Valverde, E., Julián, E., Torrents, E., Vázquez, S., (2015). Antibacterial activity of novel benzopolycyclic amines Bioorganic and Medicinal Chemistry 23, (2), 290-296

Staphylococcus aureus, especially strains resistant to multiple antibiotics, is a major pathogen for humans and animals. In this paper we have synthesized and evaluated the antibacterial activity of a new series of benzopolycyclic amines. Some of them exhibited μM MIC values against Staphylococcus aureus and other bacteria, including methicillin-resistant S. aureus MRSA. Compound 8 that displayed a good selectivity index, showed to be active in eliminating bacterial cells forming a preexisting biofilm.

Keywords: Antibacterials, Minimal biofilm inhibitory concentration, Polycyclic compounds, Staphylococcus aureus


Van Der Hofstadt, M., Hüttener, M., Juárez, A., Gomila, G., (2015). Nanoscale imaging of the growth and division of bacterial cells on planar substrates with the atomic force microscope Ultramicroscopy 154, 29-36

Abstract With the use of the atomic force microscope (AFM), the Nanomicrobiology field has advanced drastically. Due to the complexity of imaging living bacterial processes in their natural growing environments, improvements have come to a standstill. Here we show the in situ nanoscale imaging of the growth and division of single bacterial cells on planar substrates with the atomic force microscope. To achieve this, we minimized the lateral shear forces responsible for the detachment of weakly adsorbed bacteria on planar substrates with the use of the so called dynamic jumping mode with very soft cantilever probes. With this approach, gentle imaging conditions can be maintained for long periods of time, enabling the continuous imaging of the bacterial cell growth and division, even on planar substrates. Present results offer the possibility to observe living processes of untrapped bacteria weakly attached to planar substrates.

Keywords: Atomic Force Microscope (AFM), Living cell imaging, Bacteria division, Gelatine immobilization, Dynamic jumping mode


del Moral-Zamora, Beatriz, Punter-Villagrassa, Jaime, Oliva-Brañas, Ana M., Álvarez-Azpeitia, Juan Manuel, Colomer-Farrarons, Jordi, Samitier, Josep, Homs-Corbera, Antoni, Miribel-Català, Pere Ll, (2015). Combined dielectrophoretic and impedance system for on-chip controlled bacteria concentration: application to Escherichia coli Electrophoresis 36, (9-10), 1130-1141

The present paper reports a bacteria autonomous controlled concentrator prototype with a user-friendly interface for bench-top applications. It is based on a micro-fluidic lab-on-a-chip and its associated custom instrumentation, which consists in a dielectrophoretic actuator, to pre-concentrate the sample, and an impedance analyser, to measure concentrated bacteria levels. The system is composed by a single micro-fluidic chamber with interdigitated electrodes and a instrumentation with custom electronics. The prototype is supported by a real-time platform connected to a remote computer, which automatically controls the system and displays impedance data used to monitor the status of bacteria accumulation on-chip. The system automates the whole concentrating operation. Performance has been studied for controlled volumes of Escherichia coli (E. coli) samples injected into the micro-fluidic chip at constant flow rate of 10 μL/min. A media conductivity correcting protocol has been developed, as the preliminary results showed distortion of the impedance analyser measurement produced by bacterial media conductivity variations through time. With the correcting protocol, the measured impedance values were related to the quantity of bacteria concentrated with a correlation of 0.988 and a coefficient of variation of 3.1%. Feasibility of E. coli on-chip automated concentration, using the miniaturized system, has been demonstrated. Furthermore, the impedance monitoring protocol had been adjusted and optimized, to handle changes in the electrical properties of the bacteria media over time.

Keywords: Autonomous Device, Bacteria Concentrator, Dielectrophoresis, Escherichia coli, Impedance Analysis


Rodríguez-Hernández, Ana G., Muñoz-Tabares, José, Godoy-Gallardo, Maria, Juárez, Antonio, Gil, Francisco-Javier, (2013). S. sanguinis adhesion on rough titanium surfaces: Effect of culture media Materials Science and Engineering: C 33, (2), 714-720

Bacterial colonization plays a key role in dental implant failure, because they attach directly on implant surface upon implantation. Between different types of bacteria associated with the oral environment, Streptococcus sanguinis is essential in this process since it is an early colonizer. In this work the relationship between titanium surfaces modified by shot blasting treatment and S. sanguinis adhesion; have been studied in approached human mouth environment. Bacteria pre-inoculated with routinary solution were put in contact with titanium samples, shot-blasted with alumina and silicon carbide, and adhesion results were compared with those obtained when bacteria were pre-inoculated with modified artificial saliva medium and on saliva pre-coated titanium samples. Our results showed that bacterial adhesion on titanium samples was influenced by culture conditions. When S. sanguinis was inoculated in routinary culture media, colonies forming unities per square millimeter presented an increment correlated with roughness and surface energy, but separated by the type of particle used during shot-blasting treatment; whereas in modified artificial saliva only a relationship between bacteria adhered and the increment in both roughness and surface energy were observed, regardless of the particle type. Finally, on human saliva pre-coated samples no significant differences were observed among roughness, surface energy or particle.

Keywords: S. sanguinis, Bacterial adhesion, Titanium, Artificial saliva, Surface energy, Roughness


Sjoberg, B. M., Torrents, E., (2011). Shift in ribonucleotide reductase gene expression in pseudomonas aeruginosa during infection Infection and Immunity 79, (7), 2663-2669

The roles of different ribonucleotide reductases (RNRs) in bacterial pathogenesis have not been studied systematically. In this work we analyzed the importance of the different Pseudomonas aeruginosa RNRs in pathogenesis using the Drosophila melanogaster host-pathogen interaction model. P. aeruginosa codes for three different RNRs with different environmental requirements. Class II and III RNR chromosomal mutants exhibited reduced virulence in this model. Translational reporter fusions of RNR gene nrdA, nrdJ, or nrdD to the green fluorescent protein were constructed to measure the expression of each class during the infection process. Analysis of the P. aeruginosa infection by flow cytometry revealed increased expression of nrdJ and nrdD and decreased nrdA expression during the infection process. Expression of each RNR class fits with the pathogenicities of the chromosomal deletion mutants. An extended understanding of the pathogenicity and physiology of P. aeruginosa will be important for the development of novel drugs against infections in cystic fibrosis patients.

Keywords: Broad-host-range, Anaerobic growth, Drosophila-melanogaster, Bacterial biofilms, Escherichia-coli, Cystic-fibrosis, Model host, Virulence, Promoter, Vectors


Paytubi, S., Garcia, J., Juarez, A., (2011). Bacterial Hha-like proteins facilitate incorporation of horizontally transferred DNA Central European Journal of Biology 6, (6), 879-886

Horizontal gene transfer (HGT), non-hereditary transfer of genetic material between organisms, accounts for a significant proportion of the genetic variability in bacteria. In Gram negative bacteria, the nucleoid-associated protein H-NS silences unwanted expression of recently acquired foreign DNA. This, in turn, facilitates integration of the incoming genes into the regulatory networks of the recipient cell. Bacteria belonging to the family Enterobacteriaceae express an additional protein, the Hha protein that, by binding to H-NS, potentiates silencing of HGT DNA. We provide here an overview of Hha-like proteins, including their structure and function, as well as their evolutionary relationship. We finally present available information suggesting that, by expressing Hha-like proteins, bacteria such as Escherichia coli facilitate HGT incorporation and hence, the impact of HGT in their genetic diversity.

Keywords: Hha, H-NS, HGT DNA, Enterobacteria, Nucleoid-associated proteins, Enterica serovar typhimurium, Histone-like protein, h-ns, Escherichia-coli, Yersinia-enterocolitica, Salmonella-enterica


Torrents, E., Sjoberg, B. M., (2010). Antibacterial activity of radical scavengers against class Ib ribonucleotide reductase from Bacillus anthracis Biological Chemistry 391, (2-3), 229-234

Bacillus anthracis is a severe mammalian pathogen. The deoxyribonucleotides necessary for DNA replication and repair are provided via the ribonucleotide reductase (RNR) enzyme. RNR is also important for spore germination and cell proliferation upon infection. We show that the expression of B. anthracis class Ib RNR responds to the environment that the pathogen encounters upon infection. We also show that several anti-proliferative agents (radical scavengers) specifically inhibit the B. anthracis RNR. Owing to the importance of RNR in the pathogenic infection process, our results highlight a promising potential to inhibit the growth of B. anthracis early during infection.

Keywords: Anthrax, Antibacterial drug, Antibacterial target, Enzyme inhibition


Jaramillo, M. D., Torrents, E., Martinez-Duarte, R., Madou, M. J., Juarez, A., (2010). On-line separation of bacterial cells by carbon-electrode dielectrophoresis Electrophoresis 31, (17), 2921-2928

Dielectrophoresis (DEP) represents a powerful approach to manipulate and study living cells. Hitherto, several approaches have used 2-D DEP chips. With the aim to increase sample volume, in this study we used a 3-D carbon-electrode DEP chip to trap and release bacterial cells. A continuous flow was used to plug an Escherichia coli cell suspension first, to retain cells by positive DEP, and thereafter to recover them by washing with peptone water washing solution. This approach allows one not only to analyze DEP behavior of living cells within the chip, but also to further recover fractions containing DEP-trapped cells. Bacterial concentration and flow rate appeared as critical parameters influencing the separation capacity of the chip. Evidence is presented demonstrating that the setup developed in this study can be used to separate different types of bacterial cells.

Keywords: Bacteria, Carbon electrode, Dielectrophoresis, E. coli, Separation


Barreiros dos Santos, M., Sporer, C., Sanvicens, N., Pascual, N., Errachid, A., Martinez, E., Marco, M. P., Teixeira, V., Samiter, J., (2009). Detection of pathogenic Bacteria by Electrochemical Impedance Spectroscopy: Influence of the immobilization strategies on the sensor performance Procedia Chemistry 23rd Eurosensors Conference (ed. Brugger, J., Briand, D.), Elsevier Science, BV (Lausanne, Switzerland) 1, 1291-1294

Electrochemical impedance spectroscopy (EIS) is applied to detect pathogenic E. coli O157:H7 bacteria via a label free immunoassay-based detection method. Polyclonal anti-E.coli antibodies (PAb) are immobilized onto gold electrodes following two different strategies, via chemical bond formation between antibody amino groups and a carboxylic acid containing self-assembled molecular monolayer (SAM) and alternatively by linking a biotinylated anti-E. coli to Neutravidin on a mixed-SAM. Impedance spectra for sensors of both designs for increasing concentrations of E. coli are recorded in phosphate buffered saline (PBS). The Nyquist plots can be modeled with a Randle equivalent circuit, identifying the charge transfer resistance RCT as the relevant concentration dependent parameter. Sensors fabricated from both designs are able to detect very low concentration of E. coli with limits of detection as low as 10-100 cfu/ml. The influence of the different immobilization protocols on the sensor performance is evaluated in terms of sensitivity, dynamic range and resistance against nonspecific absorption.

Keywords: Bacteria detection, Biosensors, E-coli, Impedance spectroscopy


Banos, R. C., Pons, J. I., Madrid, C., Juarez, A., (2008). A global modulatory role for the Yersinia enterocolitica H-NS protein Microbiology 154, (5), 1281-1289

The H-NS protein plays a significant role in the modulation of gene expression in Gram-negative bacteria. Whereas isolation and characterization of hns mutants in Escherichia coli, Salmonella and Shigella represented critical steps to gain insight into the modulatory role of H-NS, it has hitherto not been possible to isolate hns mutants in Yersinia. The hns mutation is considered to be deleterious in this genus. To study the modulatory role of H-NS in Yersinia we circumvented hns lethality by expressing in Y. enterocolitica a truncated H-NS protein known to exhibit anti-H-NS activity in E. coli (H-NST(EPEC)). Y. enterocolitica cells expressing H-NST(EPEC) showed an altered growth rate and several differences in the protein expression pattern, including the ProV protein, which is modulated by H-NS in other enteric bacteria. To further confirm that H-NST(EPEC) expression in Yersinia can be used to demonstrate H-NS-dependent regulation in this genus, we used this approach to show that H-NS modulates expression of the YmoA protein.

Keywords: Bacterial Proteins/biosynthesis/genetics/ physiology, DNA-Binding Proteins/biosynthesis/genetics/ physiology, Electrophoresis, Gel, Two-Dimensional, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Genes, Essential, Proteome/analysis, RNA, Bacterial/biosynthesis, RNA, Messenger/biosynthesis, Reverse Transcriptase Polymerase Chain Reaction, Sequence Deletion, Yersinia enterocolitica/chemistry/genetics/growth & development/ physiology


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