Mutations in the Global Transcription Factor CRP/CAP: Insights from Experimental Evolution and Deep Sequencing

Publication date: Available online 18 June 2019Source: Computational and Structural Biotechnology JournalAuthor(s): Pernille Ott Frendorf, Ida Lauritsen, Agnieszka Sekowska, Antoine Danchin, Morten H.H. NørholmAbstractThe Escherichia coli cyclic AMP receptor protein (CRP or catabolite activator protein, CAP) provides a textbook example of bacterial transcriptional regulation and is one of the best studied transcription factors in biology. For almost five decades a large number of mutants, evolved in vivo or engineered in vitro, have shed light on the molecular structure and mechanism of CRP. Here, we review previous work, providing an overview of studies describing the isolation of CRP mutants. Furthermore, we present new data on deep sequencing of different bacterial populations that have evolved under selective pressure that strongly favors mutations in the crp locus. Our new approach identifies more than 100 new CRP mutations and paves the way for a deeper understanding of this fascinating bacterial master regulator.
Source: Computational and Structural Biotechnology Journal - Category: Biotechnology Source Type: research

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Publication date: November 2019Source: Materials Science and Engineering: C, Volume 104Author(s): T. Arun, Suresh K. Verma, Pritam Kumar Panda, R. Justin Joseyphus, Ealisha Jha, Ali Akbari-Fakhrabadi, Pranesh Sengupta, D.K. Ray, V.S. Benitha, K. Jeyasubramanyan, P.V. SatyamAbstractNanomaterial based paints are in current demand in the area of surface protective coatings due to the significant advances made to improve their antibacterial and anticorrosion characteristics. In this work, we have developed magnetic graphene oxide (MGO) paint with the incorporation of cobalt ferrite (CF) and graphene oxide (GO) along with paint...
Source: Materials Science and Engineering: C - Category: Materials Science Source Type: research
Publication date: Available online 14 July 2019Source: Biochemical Engineering JournalAuthor(s): Alvaro R. Lara, Karim E. Jaén, Olusegun Folarin, Eli Keshavarz-Moore, Jochen BüchsAbstractThe influence of oxygen limitation on pDNA yields in cultures of Escherichia coli was studied. Cultures at maximum oxygen transfer rate (OTRmax) values of 10, 14, 30 and 45 mmol L-1 h-1 and an aerobic culture at an OTRmax of 110 mmol L-1 h-1 were performed in microtiter plates (MTPs). Dissolved oxygen tension (DOT), pH, biomass and NADH fluorescence were monitored online. An E. coli strain that const...
Source: Biochemical Engineering Journal - Category: Biochemistry Source Type: research
Abstract C2 toxin produced from Clostridium botulinum serotypes C and D has a potential role in many pathophysiological mechanisms in birds and animals. It has encompassed an ADP ribosyltransferase subunit (C2I) and a translocation/binding subunit (C2II). In the present study, we intended to produce C2I mutant proteins as recombinant subunit vaccines by using glutathione-S-transferase-gene fusion system. The mutants of this study were previously evaluated from their evolutionary imprints and suggested as suitable candidates for subunit vaccines. A synthetic C2 gene was inserted in a pGEX-2T vector, cloned and expr...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
Abstract The aims of this study were to regain new epidemiology information about frequency, drug resistance rates, and typing of Campylobacter jejuni (C. jejuni) isolates, obtained from some poultry and cattle farms, slaughterhouses, and people with diarrhea. In this regard, Minimal Inhibitory Concentration (MIC) of several antibiotics and the associated antibiotic resistance genes, including tetO, tetA, cmeB, and blaOXA-61 were evaluated. The isolates were also typed, using the Fla-RFLP method. Generally, between 233 food animal samples, 80 (34.33%) C. jejuni were isolated. Moreover, 20 out of 74 (27%) human spe...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
syl D Abstract Turtoises are a great puzzle when it comes to their bacterial flora, the composition and structure of which are still unknown in details. Its component which has been best described so far is Salmonella spp., presumably due to the threat of reptile-associated salmonellosis in humans. This investigation tried to assess and characterize intestinal bacterial flora of imported tortoises found dead during quarantine. Most of the animals carried various serovars of Salmonella showing no antimicrobial resistance. Presence of multiresistant Escherichia coli was possibly a result of industrial breeding and h...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
Abstract The emergence of CTX-M-1 producing Uropathogenic Escherichia coli (UPEC) has become a serious challenge. In addition to antimicrobial resistance, a number of virulence factors have been shown. Therefore, this study was designed to determine the prevalence of O- serogroups, phylogenetic groups, exotoxin genes, and antimicrobial resistance properties of CTX-M-1- producing UPEC. A total of 248 UPEC isolates were collected. The antibiotic resistance was performed, and PCR was used to detect the blaCTX-M1, exotoxins, serogroups and phylogroups of UPEC. Of 248 isolates, 95 (38.3%) harbored blaCTX-M-1. Of them, ...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
Abstract Unawareness of canine parasitic diseases among at-risk hosts and an uncontrolled program of stray dog population have caused that zoonotic parasites received great attention in endemic regions of the Middle East. A total of 552 faecal samples were collected between December 2016 to January 2018 from stray (n = 408) and domestic (n = 144) dogs of Iran. All specimens were coproscopically observed following concentration and flotation techniques. Subsequently, the DNAs of taeniid eggs were extracted, amplified, and sequenced by targeting of mitochondrial cytochrome oxidase subunit...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
Abstract S. Choleraesuis (Choleraesuis) and S. Typhimurium (Typhimurium) cause salmonellosis in pigs and humans. The effects of vaccine strains pSV-less Typhimurium OU5048 and Choleraesuis OU7266 and SPI-2-mutant Choleraesuis SC2284 on the immune responses of pigs against Typhimurium, Choleraesuis, and S. Enteritidis (Enteritidis) with or without the virulence plasmid (pSV) were determined. After oral vaccination of three vaccine groups and challenge with Choleraesuis CN36, the level of Salmonella-specific IgG in sera and the bactericidal effects and superoxide generation of peripheral blood mononuclear cells (PBM...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
In this study the effects against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Acinetobacterbaumannii, Streptococcus pyogenes, Proteus vulgaris, Salmonella typhi, Shigella spp., Candida albicans, Candida parapsilosis, Anopheles stephensi, Aedes aegypti, A. aegypti, A. albopictus, Culex pipiens, Trypanosoma brucei, Leishmania major, Trichomonas. Vaginalis, poliovirus type 1, coxsackie virus B, echovirus 6, West Nile Virus, herpes virus type 1, HSV-1 virus, Fusarium spp., Aspergillus flavus, Aspergillus niger, C. albicans, Alternaria alternata, Setosphaeria turcica and Magnaporthe grisea was revealed. ...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
In this study, a new transaminase from Pseudomonas fluorescens ZJB09-108 (named as Pf-TA) was cloned and expressed in E. coli BL21 (DE3). The amino acid sequence analysis showed that the maximum sequence identity of Pf-TA with other transaminases is 74%. This recombinant Pf-TA exhibited a good temperature stability and pH stability. The Km and kcat of Pf-TA toward PPO were 34.02 mmol·L-1 and 11.3 s-1, respectively. After optimization of the L-PPT biosynthesis catalyzed by whole-cells of E. coli harboring transaminase,>99% yield of L-PPT with 99.9% of enantiomeric excess was attained at a PPO concentration ...
Source: Process Biochemistry - Category: Biochemistry Source Type: research
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