• Title/Summary/Keyword: bacterial expression

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A Study on Microbial Aspects of Korean Human Milk by Collection Methods (수집방법에 따른 한국인 모유의 미생물 분포에 관한 연구)

  • 이조윤;배형철;남명수
    • Food Science of Animal Resources
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    • v.23 no.3
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    • pp.269-277
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    • 2003
  • This study was carried out to evaluate the safety of Korean human milk. The microorganisms were identified from human milk of 149 healthy mothers by two collection methods, hand and pump expression. The means of total bacterial counts were 2.33x10$^4$ cfu/mL on the samples collected by the pump expression and 7.83xl0$^3$ cfu/mL on those collected by the hand expression. Therefore, the total bacterial counts of pump expression samples was 9.80xl0$^2$∼3.06x10$^4$ cfu/mL more than that of hand expression samples. The coliform counts of pump expression was 9.36xl0$^3$∼8.57xl0$^4$ cfu/mL more than that of hand expression. However, there was any significant differences of the lactic acid bacterial counts between the two samples collected by each methods. 100 strains of 5 patterns of total bacterial counts were isolated based on the morphology of colony in the standard plate count agar. 13 species were identified among the isolated strains. The dominant species in Korean human milk were Staphylococcus which 7 subspecies identified(81% in the rate of total bacteria, 1.07x10$^4$ cfu/mL). Other species identified were Micrococcus, Bacillus, Providencia, Pseudomonas, Yersinia and Acinetobacter. 36 strains of 6 patterns of lactic acid bacterial counts were isolated based on morphology of colony in the BCP agar. 7 species were identified among the isolated strains. The dominant species of lactic acid bacteria in Korean human milk were Lactobacillus brevis(50.9% in the rate of lactic acid bacteria, 4.72xl0$^4$ cfu/mL). Others species identified(49.1% lactic acid bacteria) were Lactobacillus curvatus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus acidophilus, Leuconostic lactis and Streptococcus salivarius subsp. thermophilus.

Stimulation of Platelet-Activating Factor (PAF) Synthesis in Human Intestinal Epithelial Cell Line by Aerolysin from Aeromonas encheleia

  • Nam In-Young;Cho Jae-Chang;Myung Hee-Joon;Joh Ki-Seong
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1292-1300
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    • 2006
  • Aeromonas encheleia, a potential human intestinal pathogen, was shown to infect a human intestinal epithelial cell line (Caco-2) in a noninvasive manner. The transcriptional profile of the Caco-2 cells after infection with the bacteria revealed an upregulated expression of genes involved in chloride secretion, including that of phospholipase A2 (PLA2) and platelet-activating factor (PAF) acetylhydrolase (PAFAH2). This was also confirmed by a real-time RT-PCR analysis. As expected from PLA2 induction, PAF was produced when the Caco-2 cells were infected with the bacteria, and PAF was also produced when the cells were treated with a bacterial culture supernatant including bacterial extracellular proteins, yet lacking lipopolysaccharides. Bacterial aerolysin was shown to induce the production of PAF.

Bacterial Quorum Sensing and Anti-Quorum Sensing (세균의 적정밀도 인식을 통한 신호전달 및 신호전달 차단 연구)

  • 박순양;이정기
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.1-10
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    • 2004
  • Many bacteria monitor their population density and control the expression of specialized gene sets in response to bacterial cell density based on a mechanism referred to as quorum sensing. In all cases, quorum sensing involves the production and detection of extracellular signaling molecules, auto inducers, as which Gram-negative and Gram-positive bacteria use most prevalently acylated homoserine lactones and processed oligo-peptides, respectively. Through quorum-sensing communication circuits, bacteria regulate a diverse array of physiological functions, including virulence, symbiosis, competence, conjugation, antibiotic production, motility, sporulation, and biofilm formation. Many pathogens have evolved quorum-sensing mechanisms to mount population-density-dependent attacks to over-whelm the defense responses of plants, animals, and humans. Since these AHL-mediated signaling mechanisms are widespread and highly conserved in many pathogenic bacteria, the disruption of quorum-sensing system might be an attractive target for novel anti-infective therapy. To control AHL-mediated pathogenicity, several promising strategies to disrupt bacterial quorum sensing have been reported, and several chemicals and enzymes have been also investigated for years. These studies indicate that anti-quorum sensing strategies could be developed as possible alternatives of antibiotics.

Relation of Plant Age to Bacterial Multiplication in Pepper and Tomato Leaves Inoculated with Xanthomonas campestris pv. vesicatoria (Xanthomonas campestris pv. veicatoria에 감염된 고추와 토마토잎에서의 세규증식과 식물나이와의관계)

  • 이종탁;황병국
    • Korean Journal Plant Pathology
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    • v.10 no.1
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    • pp.18-24
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    • 1994
  • Multiplications and pathogenic reactions of different pepper and tomato strains of Xanthomonas campestris pv. vesicatoria were evaluated in the most upper leaves of pepper and tomato plants at different growth stages. Hypersensitive reactions were induced in mature pepper plants by inoculation with only the tomato strains but not with the pepper strains, suggesting the expression of age-related resistance in pepper plants. The age-related resistance also seems to be correlated with an apparent inability of the bacteria to multiply as extensively in mature as in young plants. No significant differences among the Korean and U. S. pepper cultivars tested were found in bacterial multiplication, irrespective of bacterial stain or plant growth stage. Korean tomato cultivars tested also were highly susceptible to either tomato or pepper strains during the development of tomato plants.

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Quorum Sensing Regulation of Biofilm Formation by Periodontal Pathogens

  • Choi, Bong-Kyu
    • International Journal of Oral Biology
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    • v.43 no.4
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    • pp.171-175
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    • 2018
  • Quorum sensing (QS) is a cell density-dependent communication mechanism between bacteria through small signaling molecules. When the number of QS signaling molecules reaches a threshold, they are transported back into the cells or recognized by membrane-bound receptors, triggering gene expression which affects various phenotypes including bioluminescence, virulence, adhesion, and biofilm formation. These phenotypes are beneficial for bacterial survival in harsh environments. This review summarizes the application of QS inhibitors for control of biofilm formation and virulence expression of periodontal pathogens.

Identification and Cloning of a Fraction 1 Protein of Yersinia pestis that Produces Protective Immune Responses

  • Kim Jong-Hyun;Cho Seung-Hak;Jang Hyun-Chul;Lee Hee-Cheul;Kim Young-Il;Kang Yeon-Ho;Lee Bok-Kwon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1180-1184
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    • 2006
  • The capsule that surrounds Yersinia pestis cells is composed of a protein-polysacchride complex; the purified protein component is fraction I (F1) antigen. We report the cloning of the cafl gene and its expression in Escherichia coli using the vector pETl02/D-TOPO and the F1-specific monoclonal antibody. The recombinant F1 (rF1) antigen had a molecular size of 17.5 kDa, which was identical to that of the F1 antigen produced by Y. pestis. Recombinant F1 protein was found to react to polyclonal antiserum to Y. pestis Fl. Recombinant F1 was purified by ProBond purification system and induced a protective immune response in BALB/c mice challenged with up to 10$^5$ virulent Y. pestis. Purified rF1 protein was used in an ELISA to evaluate the ability of a method to detect antibodies to Y. pestis in animal sera. These results strongly indicated that the rF1 protein is a suitable species-specific immunodiagnostic antigen and vaccine candidate.

Expression of Antimicrobial Peptide (AMP), Moricin Using SUMO Fusion Tag in Escherichia coli (대장균에서 SUMO fusion tag을 이용하여 항균펩타이드인 moricin의 발현)

  • Ahn, Dong-gyu;Park, Sun Ill;Kim, Soon Young
    • Journal of Life Science
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    • v.32 no.12
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    • pp.956-961
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    • 2022
  • Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.

DNA Microarray and Gene Ontology Enrichment Analysis Reveals That a Mutation in opsX Affects Virulence and Chemotaxis in Xanthomonas oryzae pv. oryzae

  • Kim, Hong-Il;Park, Young-Jin
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.190-200
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    • 2016
  • Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial leaf blight (BLB) in rice (Oryza sativa L.). In this study, we investigated the effect of a mutation in opsX (XOO1056), which encodes a saccharide biosynthesis regulatory protein, on the virulence and bacterial chemotaxis of Xoo. We performed DNA micro-array analysis, which showed that 63 of 2,678 genes, including genes related to bacterial motility (flagellar and chemotaxis proteins) were significantly downregulated ($<\;-2\;log_2$ fold changes) by the mutation in opsX. Indeed, motility assays showed that the mutant strain was nonmotile on semisolid agar swarm plates. In addition, a mutant strain (opsX::Tn5) showed decreased virulence against the susceptible rice cultivar, IR24. Quantitative real-time RT-PCR reaction was performed to confirm the expression levels of these genes, including those related to flagella and chemotaxis, in the opsX mutant. Our findings revealed that mutation of opsX affects both virulence and bacterial motility. These results will help to improve our understanding of Xoo and provide insight into Xoo-rice interactions.

Acyl Homoserine Lactone in Interspecies Bacterial Signaling

  • Kanojiya, Poonam;Banerji, Rajashri;Saroj, Sunil D.
    • Microbiology and Biotechnology Letters
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    • v.50 no.1
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    • pp.1-14
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    • 2022
  • Bacteria communicate with each other through an intricate communication mechanism known as quorum sensing (QS). QS regulates different behavioral aspects in bacteria, such as biofilm formation, sporulation, virulence gene expression, antibiotic production, and bioluminescence. Several different chemical signals and signal detection systems play vital roles in promoting highly efficient intra- and interspecies communication. Gram-negative bacteria coordinate gene regulation through the production of acyl homoserine lactones (AHLs). Gram-positive bacteria do not code for AHL production, while some gram-negative bacteria have an incomplete AHL-QS system. Despite this fact, these microbes can detect AHLs owing to the presence of LuxR solo receptors. Various studies have reported the role of AHLs in interspecies signaling. Moreover, as bacteria live in a polymicrobial community, the production of extracellular compounds to compete for resources is imperative. Thus, AHL-mediated signaling and inhibition are considered to affect virulence in bacteria. In the current review, we focus on the synthesis and regulation mechanisms of AHLs and highlight their role in interspecies bacterial signaling. Exploring interspecies bacterial signaling will further help us understand host-pathogen interactions, thereby contributing to the development of therapeutic strategies intended to target chronic polymicrobial infections.

Cooperative Interaction between Acid and Copper Resistance in Escherichia coli

  • Kim, Yeeun;Lee, Seohyeon;Park, Kyungah;Yoon, Hyunjin
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.602-611
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    • 2022
  • The persistence of pathogenic Escherichia coli under acidic conditions poses a serious risk to food safety, especially in acidic foods such as kimchi. To identify the bacterial factors required for acid resistance, transcriptomic analysis was conducted on an acid-resistant enterotoxigenic E. coli strain and the genes with significant changes in their expression under acidic pH were selected as putative resistance factors against acid stress. These genes included those associated with a glutamate-dependent acid resistance (GDAR) system and copper resistance. E. coli strains lacking GadA, GadB, or YbaST, the components of the GDAR system, exhibited significantly attenuated growth and survival under acidic stress conditions. Accordantly, the inhibition of the GDAR system by 3-mercaptopropionic acid and aminooxyacetic acid abolished bacterial adaptation and survival under acidic conditions, indicating the indispensable role of a GDAR system in acid resistance. Intriguingly, the lack of cueR encoding a transcriptional regulator for copper resistance genes markedly impaired bacterial resistance to acid stress as well as copper. Conversely, the absence of YbaST severely compromised bacterial resistance against copper, suggesting an interplay between acid and copper resistance. These results suggest that a GDAR system can be a promising target for developing control measures to prevent E. coli resistance to acid and copper treatments.