• Title/Summary/Keyword: Virulent strain

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Identification of Differentially Displayed Genes of a Pseudomonas Resistant Soybean (Glycine max)

  • Kang, Sang-Gu;Cha, Hyeon-Wook;Chang, Moo-Dng;Park, Eui-Ho
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.239-247
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    • 2003
  • In Korea, a local soybean (Glycine max) genotype 56l. was found to be strongly resistant to a virulent bacterial strain of a Pseudomonas sp. SN239. Specific genes involved in the resistance of the soybean genotype 561 were identified and the pattern of gene expression against the Pseudomonas infection was analyzed using differential-display reverse transcription PCR (DDRT-PCR). More than 126 cDNA fragments representing mRNAs were induced within 48 hours of bacteria inoculation. Among them, 28 cDNA fragments were cloned and sequenced. Twelve differentially displayed clones with open reading frames had unknown functions. Sixteen selected cDNA clones were homologous to known genes of other organisms. Some of the identified cDNAs were pathogenesis-related (PR) genes and PR-like genes. These cDNAs included a putative calmodulin-binding protein; an endo-l,3-1,4-$\bate$-D-glucanase; a $\bate$-1,3-endoglucanase; a $\bate$-1,3-exoglucanase; a phytochelatin synthetase-like gene; a thiol protease; a cycloartenol synthase; and a putative receptor-like serine/threonine protein kinase. Among them, four genes were found to be putative PR genes induced significantly by the Pseudomonas infection. These included a calmodulin-binding protein gene, a $\bate$-1,3-endoglucanase gene, a receptor-like serine/threonine protein kinase gene, and pS321 (unknown function). These results suggest that the differentially expressed genes may mediate the strong resistance of soybean 561 to the strain SN239 of Pseudomonas sp.

Selection of Antagonistic Bacteria for Biocontrol of Botrytis cinerea Causing Gray Mold on Vitis spp (포도 잿빛곰팡이병의 생물적 방제를 위한 길항세균 선발)

  • Seo, Sang-Tae;Park, Jong-Han;Han, Kyoung-Suk;Cheong, Seung-Ryong
    • Research in Plant Disease
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    • v.12 no.3
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    • pp.267-271
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    • 2006
  • Botrytis cinerea Pers. was found to be highly virulent to the grapevine plant, especially in greenhouse condition. Pseudomonas species play key roles for the biocontrol of many plant diseases especially in soil. Of the 83 isolates of Pseudomonas spp., a bacterial strain P84, isolated from tomato rhizosphere, was shown to suppress a wide range of phytopathogenic fungi in vitro. The isolate was identified as Pseudomonas putida on the basis of its bacteriological and genetic characteristics. The P. putida P84 strain carry the phlD gene for 2,4-diacetylphloroglucinol biosynthesis and may produce the antibiotics as an antagonistic mechanism involved in biocontrol. The antagonistic activity of the bacterium has a promising implication for its use as a biocontrol agent to control grapevine gray mold.

Role of TolC in Vibrio vulnificus Virulence in Mice

  • Lin Mei-Wei;Lin Chen-Hsing;Tsai Shih-Feng;Hor Lien-I
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.59-62
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    • 2002
  • The role of a TolC homologue in the virulence of Vibrio vulnificus, a marine bacterium causing serious wound infection and fulminant septicemia in persons with underlying conditions, has been studied. TolC, an outer membrane protein, has been implicated in a variety of bacterial functions including export of diverse molecules ranging from large proteins to antibiotics. A homologue of the tolC gene of V. cholerae, which has been shown to be required for bile resistance, cytotoxicity and colonization of this organism, was identified in the partially determined genome sequence of V. vulnificus. To determine the role of TolC in the virulence of V. vulnificus, a TolC-deficient (TD) mutant was isolated by in vivo allelic exchange. Compared with the parent strain, the TD mutant was more sensitive to bile, and much less virulent in mice challenged subcutaneously. This mutant was noncytotoxic to the HEp-2 cells, but its metalloprotease and cytolysin activities in the culture supernatant were comparable to the parent strain. In addition, the resistance of the TD mutant to human serum bactericidal activity as well as its growth in either human or murine blood was not affected. Collectively, our data suggest that TolC may be involved in colonization and/or spread of V. vulnificus to the blood stream, probably by secreting a cytotoxin other than the cytolysin.

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Genetic characteristics of the Korean isolate KI -1 of Toxoplasma gondii

  • LIN Aifen;SHIN Eun-Hee;KIM Tae-Yun;PARK Jae-Hwan;GUK Sang-Mee;CHAI Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.43 no.1 s.133
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    • pp.27-32
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    • 2005
  • Toxoplasma gondii tachyzoites were isolated from an ocular patient in the Republic of Korea and maintained in the laboratory (designated KI-1). In the present study, its genotype was determined by analyzing dense granule antigen 6 (GRA6) gene and surface antigen 2 (SAG2) gene as typing markers. Digestion of the amplification products of GRA6 and of the 5' and 3' ends of SAG2, respectively, with Mse I, Sau3A I, and Hha I, revealed that KI-1 is included in the genotype I, which includes the worldwide virulent RH strain. In addition, when the whole sequences of the coding regions of SAG1, rhoptry antigen 1 (ROP1), and GRA8 genes of KI-1 were compared with those of RH, minor nucleotide polymorphisms and amino acid substitutions were identified. These results show that KI-1 is a new geographical strain of T. gondii that can be included in the genotype I.

Biological Control using Bacillus toyonensis Strain CAB12243-2 against Soft Rot on Chinese Cabbage (Bacillus toyonensis CAB12243-2 균주를 이용한 배추 무름병의 생물적 방제)

  • Kim, Byung-Ryun;Park, Myung-Soo;Han, Kwang-Seop;Hahm, Soo-Sang;Park, In-Hee;Song, Jae-Kyeong
    • Korean Journal of Organic Agriculture
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    • v.26 no.1
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    • pp.129-140
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    • 2018
  • Pectobacterium carotovorum subsp. carotovorum was found to be highly virulent to various vegetables, including Chinese cabbage. The antibacterial isolate CAB12243-2 was tested in a field bioassay for suppressing soft rot disease. The nucleotide sequencing of the 16S rRNA gene identified, the CAB12243-2 strain used in this study as Bacillus toyonensis. B. toyonensis CAB12243-2 inhibited the pectate lyase process by soft rot pathogens, and used trehalose and glucose as carbon sources. In field tests, the antibacterial isolate B. toyonensis CAB12243-2 suppressed soft rot disease with 73.0% control efficacy on the spring cultivar "Norangbom" and with 68.9% efficacy on the fall cultivar "Bulam 3". These results suggest that B. toyonensis CAB12243-2 can be used as a biological control agent for the control of soft rot diseases on vegetables.

Salmonella vector induces protective immunity against Lawsonia and Salmonella in murine model using prokaryotic expression system

  • Sungwoo Park;Eunseok Cho;Amal Senevirathne;Hak-Jae Chung;Seungmin Ha;Chae-Hyun Kim;Seogjin Kang;John Hwa Lee
    • Journal of Veterinary Science
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    • v.25 no.1
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    • pp.4.1-4.14
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    • 2024
  • Background: Lawsonia intracellularis is the causative agent of proliferative enteropathy and is associated with several outbreaks, causing substantial economic loss to the porcine industry. Objectives: In this study, we focused on demonstrating the protective effect in the mouse model through the immunological bases of two vaccine strains against porcine proliferative enteritis. Methods: We used live-attenuated Salmonella Typhimurium (ST) secreting two selected immunogenic LI antigens (Lawsonia autotransporter A epitopes and flagellin [FliC]-peptidoglycan-associated lipoprotein-FliC) as the vaccine carrier. The constructs were cloned into a Salmonella expression vector (pJHL65) and transformed into the ST strain (JOL912). The expression of immunogenic proteins within Salmonella was evaluated via immunoblotting. Results: Immunizing BALB/c mice orally and subcutaneously induced high levels of LI-specific systemic immunoglobulin G and mucosal secretory immunoglobulin A. In immunized mice, there was significant upregulation of interferon-γ and interleukin-4 cytokine mRNA and an increase in the subpopulations of cluster of differentiation (CD) 4+ and CD 8+ T lymphocytes upon splenocytes re-stimulation with LI antigens. We observed significant protection in C57BL/6 mice against challenge with 106.9 times the median tissue culture infectious dose of LI or 2 × 109 colony-forming units of the virulent ST strain. Immunizing mice with either individual vaccine strains or co-mixture inhibited bacterial proliferation, with a marked reduction in the percentage of mice shedding Lawsonia in their feces. Conclusions: Salmonella-mediated LI gene delivery induces robust humoral and cellular immune reactions, leading to significant protection against LI and salmonellosis.

Characterization of the genomes of Aujeszky's disease virus isolated in Korea (국내분리 오제스키병 바이러스의 게놈 유전자 특성 분석)

  • Hyun, Bang-Hun;Kim, In-Joong;Pyo, Hyun-Mi;Cha, Sang-Ho;Park, Ji-Yeun;Song, Jae-Young;Cho, In-Soo;Yang, Chang-Bum;An, Soo-Hwan;Lee, Joong-Bok
    • Korean Journal of Veterinary Research
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    • v.49 no.1
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    • pp.45-57
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    • 2009
  • The molecular genetic characterization of Aujeszky's disease virus (ADV) Yangsan strain (ADVYS), a Korean isolate, was investigated by analyzing the electrophoresis patterns and the physical maps of the viral DNA digested with various endonucleases. To establish DNA library for ADV-YS, twelve major BamHI restricted segments were cloned. Each location of the segments in the ADV genome was determined by sequence comparison with the sequences reported in Genbank and those sequences of the both termini of the segments. Physical maps were constructed based on the electrophoresis patterns of the digested viral DNA by restriction endonuclease and the results of Southern blot analyses with various DIG labeled probes originated from those of enzyme restricted segments of virulent (Shope) and avirulent (Bartha) strain. Comparing ADV-YS with a standard strain of Kaplan in the maps of restriction enzymes, following major respects were identified: (i) disappearance of BamHI restriction site between the first and second BamHI segments, (ii) creation of the BamHI restriction site in the fifth segment, and (iii) generation of the BglII site in the unique short (US) region. The genome of ADV-YS also contains a type 2 herpesvirus DNA molecule (in which the US region only inverts itself relative to the unique longregion) like all other ADV strains except Norden strain(type3), analyzed up to date. The size of the ADV genome estimated from the sizes of the restriction enzyme fragments, was approximately 145.3 kb (BamHI) or 145.4 kb (BglII). BamHI enzyme cleavage patterns were compared among the five Korean ADV isolates: Yangsan, Yongin, Dangjin, Jincheon and Iksan strains. Difference either in the number or in the size of the DNA fragments, suspected regions of termini of IR and TR, could be detected among all five strains.

Protective Immunity Induced by Systemic and Mucosal Delivery of DNA Vaccine Expressing Glycoprotein B of Pseudorabies Virus

  • Yoon, Hyun-A;Han, Young-Woo;Aleyas, Abi George;George, June Abi;Kim, Seon-Ju;Kim, Hye-Kyung;Song, Hee-Jong;Cho, Jeong-Gon;Eo, Seong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.591-599
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    • 2008
  • A murine model immunized by systemic and mucosal delivery of plasmid DNA vaccine expressing glycoprotein B (pCIgB) of pseudorabies virus (PrV) was used to evaluate both the nature of the induced immunity and protection against a virulent virus. With regard to systemic delivery, the intramuscular (i.m.) immunization with pCIgB induced strong PrV-specific IgG responses in serum but was inefficient in generating a mucosal IgA response. Mucosal delivery through intranasal (i.n.) immunization of pCIgB induced both systemic and mucosal immunity at the distal mucosal site. However, the levels of systemic immunity induced by i.n. immunization were less than those induced by i.m. immunization. Moreover, i.n. genetic transfer of pCIgB appeared to induce Th2-biased immunity compared with systemic delivery, as judged by the ratio of PrV-specific IgG isotypes and Th1- and Th2-type cytokines produced by stimulated T cells. Moreover, the immunity induced by i.n. immunization did not provide effective protection against i.n. challenge of a virulent PrV strain, whereas i.m. immunization produced resistance to viral infection. Therefore, although i.n. immunization was a useful route for inducing mucosal immunity at the virus entry site, i.n. immunization did not provide effective protection against the lethal infection of PrV.

Comparison of Virulence Factors, Phylogenetic Groups and Ciprofloxacin Susceptibility of Escherichia coli Isolated from Healthy Students and Patients with Urinary Tract Infections in Korea

  • Park, Min;Park, Soon-Deok;Kim, Sa-Hyun;Woo, Hyun-Jun;Lee, Gyu-Sang;Kim, Hyun-Woo;Yang, Ji-Young;Cho, Eun-Hee;Uh, Young;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.146-151
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    • 2012
  • Urinary tract infection (UTI) is one of the most common bacterial infections and is predominantly caused by uropathogenic Escherichia coli (UPEC). UPEC strains generally possess several genes encoding virulent factors, which are mostly adhesins, toxins, bacteriocin and siderophores. E. coli is composed of four main phylogenetic group (A, B1, B2, D) and virulent extra-intestinal strains mainly belong to groups B2 and D. Prescription of ciprofloxacin, a kind of fluoroquinolone group antibiotics, is increasing now a days, but resistance to this drug is also increasing. A total of 188 strains of E. coli were collected. Thirteen strains were collected from healthy students in 2011 and 175 strains from patients with urinary tract infection in 2010. Virulence factor genes (papC, fimG/H, sfaD/E, hlyA, cnf1, and usp) were amplified by polymerase chain reaction (PCR) methods for phylogenetic group (A, B1, B2, D) detection. Ciprofloxacin susceptibility test was performed by disk diffusion method. The identified virulence factors (VFs), phylogenetic groups and ciprofloxacin resistance in 13 E. coli strains isolated from healthy students were papC (15.4%), fimG/H (76.9%), sfaD/E (30.8%), hlyA (23.1%), cnf1 (23.1%), usp (7.7%), phylogenetic group A (23%), B1 (8%), B2 (46%), D (23%) and ciprofloxacin resistance (7.7%), while those of in 175 E. coli strains isolated from patients with UTI were papC (41.1%), fimG/H (92.5%), sfaD/E (30.3%), hlyA (10.3%), cnf1 (30.3%), usp (27.4%), phylogenetic group A (9.1%), B1 (5.1%), B2 (60.6%), D (25.1%) and ciprofloxacin resistance (29.7%). In this study, 10 out of 13 E. coli strains (76.9%) from healthy students were found to possess more than one virulence factor associated with adhesion. In addition, one E. coli strain isolated from healthy students who had never been infected with UPEC showed ciprofloxacin resistance. According to these results between the virulence factors and phylogenetic groups it was closely associated, and UPEC strains isolated from patients showed high level of ciprofloxacin resistance.

Studies on the safety of Brucella abortus RB51 vaccine I. Comparison of the biochemical and genetic characteristics of Brucella abortus RB51 vaccine strains (부루세라백신(RB51)의 안전성에 관한 연구 I. Brucella abortus RB51 백신균주의 생화학적 및 유전학적 성상비교)

  • Kim, Jong-man;Woo, Sung-ryong;Lee, Ji-youn;Jung, Suk-chan;Kang, Seung-won;Kim, Jong-yeom;Yoon, Yong-dhuk;Cho, Sang-nae;Yoo, Han-sang;Olsen, Steven C.
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.533-541
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    • 2000
  • Biochemical and genetic analysis were carried out to investigate the potential recovery of pathogenecity or related mutations of Brucella abortus RB51 vaccine strains. RB51 strains were recovered from commercial vaccines, including related seed stocks from private companies in Republic of Korea, strain from USA, a reference strain from C university and a field isolate (Daehungjin) from aborted dairy cow after RB51 vaccination were compared with two identified virulent wild strains (S2308 and a field strain isolated from dairy cow in Korea) at the same conditions. All the strains examined, except identified pathogenic strains, revealed the identical characteristics to the original RB51 in biochemical properties, antigen and bacteriophage typing. Outer membrane protein (OMP) profiles from strains of RB51 showed the same patterns with standard RB51 in SDS-PAGE. In addition, Western blotting with the brucella specific monoclonal antibody also indicated that all the vaccine strains were completely deficient in their LPS compared to the pathogenic Br abortus strains. The differences in DNA structures among strains were also possible to detect after PCR. All vaccine strains, except S19, S1119-3, S1075, S544 and Br suis, were amplified a 178bp DNA fragment of eri-gene, and 364bp of IS711 elements. In contrast, 498bp DNA product was only found with Br abortus. Overall evidences in the present study confirmed that the RB51 strains for vaccine production in Korea did not originated from the phenomena of possible recovery of pathogenicity or related to any potential mutation event at all.

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