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Keywords = Pil fimbriae

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17 pages, 3611 KiB  
Article
Recombinant PilS: Cloning, Expression and Biochemical Characterization of a Pil-Fimbriae Subunit
by Danielle D. Munhoz, Jessika C. A. Silva, Natalia C. Freitas, Leo K. Iwai, Karina A. Aires, Christiane Y. Ozaki, Cristiane S. Souza, Letícia B. Rocha, Miriam A. Silva, Izabella M. Henrique, Waldir P. Elias, Eneas Carvalho, Ligia Morganti, Rosa M. Chura-Chambi and Roxane M. F. Piazza
Microorganisms 2022, 10(6), 1174; https://doi.org/10.3390/microorganisms10061174 - 7 Jun 2022
Viewed by 2426
Abstract
Pil-fimbriae is a type IV pili member, which is a remarkably versatile component with a wide variety of functions, including motility, attachment to different surfaces, electrical conductance, DNA acquisition, and secretion of a broad range of structurally distinct protein substrates. Despite the previous [...] Read more.
Pil-fimbriae is a type IV pili member, which is a remarkably versatile component with a wide variety of functions, including motility, attachment to different surfaces, electrical conductance, DNA acquisition, and secretion of a broad range of structurally distinct protein substrates. Despite the previous functional characterization of Pil, more studies are required to understand the regulation of Pil expression and production, since the exact mechanisms involved in these steps are still unknown. Therefore it is extremely important to have a protein with the correct secondary and tertiary structure that will enable an accurate characterization and a specific antisera generation. For this reason, the aim of this work was to generate potential tools for further investigations to comprehend the mechanisms involved in Pil regulation and its role in pathogenic E. coli infections with the obtaining of a precise native-like recombinant PilS and the corresponding antisera. The pilS gene was successfully cloned into an expression vector, and recombinant PilS (rPilS) was efficiently solubilized and purified by metal affinity chromatography. Protein characterization analyses indicated that rPilS presented native-like secondary and tertiary structures after the refolding process. The generated anti-rPilS sera efficiently recognized recombinant and native proteins from atypical enteropathogenic E. coli strains. Full article
(This article belongs to the Section Molecular Microbiology and Immunology)
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14 pages, 1306 KiB  
Article
Prevalence of the Genes Associated with Biofilm and Toxins Synthesis amongst the Pseudomonas aeruginosa Clinical Strains
by Tomasz Bogiel, Dagmara Depka, Mateusz Rzepka, Joanna Kwiecińska-Piróg and Eugenia Gospodarek-Komkowska
Antibiotics 2021, 10(3), 241; https://doi.org/10.3390/antibiotics10030241 - 28 Feb 2021
Cited by 25 | Viewed by 4079
Abstract
Pseudomonas aeruginosa is one of the most commonly isolated bacteria from clinical specimens, with an increasing isolation frequency in nosocomial outbreaks. The hypothesis tested was whether carbapenem-resistant P. aeruginosa strains display an altered carriage of the virulence factor genes, depending on the type [...] Read more.
Pseudomonas aeruginosa is one of the most commonly isolated bacteria from clinical specimens, with an increasing isolation frequency in nosocomial outbreaks. The hypothesis tested was whether carbapenem-resistant P. aeruginosa strains display an altered carriage of the virulence factor genes, depending on the type of carbapenem resistance. The aim of the study was to investigate, by PCR, the frequency of 10 chosen virulence factors genes (phzM, phzS, exoT, exoY, exoU, toxA, exoS, algD, pilA and pilB) and the genotype distribution in 107 non-duplicated carbapenem-resistant P. aeruginosa isolates. P. aeruginosa genes involved in phenazine dyes and exoenzyme T synthesis were noted with the highest frequency (100%). Fimbriae-encoding genes were detected with the lowest incidence: 15.9% and 4.7% for pilin A and B, respectively. The differences observed between the exoS gene prevalence amongst the carbapenemase-positive and the carbapenemase-negative strains and the pilA gene prevalence amongst the strains of different origins were statistically significant. Virulence genes’ prevalence and the genotype distribution vary amongst P. aeruginosa strains resistant to carbapenems, especially in terms of their carbapenemase synthesis ability and the strain origin. Full article
(This article belongs to the Special Issue The Biofilm Hurdle; Diagnostics and Treatment Challenges, Resistance Development, Gene Exchange, Evolution and Identification)
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