• Title/Summary/Keyword: E. coli(K99)

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Effect of autogenous Escherichia coli vaccine in pig (돼지에서 대장균 자가백신 효과)

  • 윤교복;김종술;정동수;박양주;이유섭;한정희
    • Korean Journal of Veterinary Service
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    • v.21 no.2
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    • pp.117-126
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    • 1998
  • This study was performed to investigate the immunogenicity of autogenous E coli vaccines and their preventive effects on diarrhea in suckling piglets. Autogenous E coli live and killed vaccines were made from the E coli strains isolated from piglets showing diarrhea in field. In group I, pregnant sows were administered with live and killed vaccines at 4 and 2 weeks before parturition, respectively, Killed vaccines were administered twice to pregnant sows at 4 and 2 weeks before parturition in group II, and saline instead of autogenous E coli vaccines was administered to pregnant sows in group III for the control. After parturition, antibody titers in colostrum and milk from sows, incidence of diarrhea in suckling piglets, and immunoreactivity in the ileum of piglets from each treatment group were examined. The results were as follows ; 1. Sixty-two strains of E coli were isolated from suckling piglets with diarrhea. Of the strains, K88 pilus and K99 pilus antigens were identified in 6(9.8%) and 4(6.5%), respectively. Molecular weights of K88 and K99 pilus were 27,500 and 18,500 daltons, respectively. 2. Antibody titers in colostrum from sows after parturition were 1 : 512 to 1 : 1,024 in group I, 1.256 to 1.512 in group II, and 1 : 4 to 1 : 16 in group III. 3. The incidences of diarrhea In suckling piglets of group I, II and III were 3.3%, 9.4% and 21.4%, respectively. 4. When the immunoreactivity in the ileum of piglets from each group was examined, the proportion of IgG-immunoreactivity cells in group I or II was higher than that in group III. In conclusion, administration of autogenous E coli vaccines to pregnant sows before parturition can be an effective way to prevent diarrhea in suckling piglets.

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Transformation of Edwardsiella tarda and Transcriptional Characteristics of E-lysis Gene in Recombinant Bacterial Ghosts (어류 병원성 세균 Edwardsiella tarda의 형질전환 및 재조합 ghost 세균에서의 E-lysis 유전자의 전사 발현 특징)

  • Kwon, Se Ryun;Nam, Yoon Kwon
    • Korean Journal of Ichthyology
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    • v.19 no.2
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    • pp.83-87
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    • 2007
  • Edwardsiella tarda, a gram (-) pathogen causing edwardsiellosis in farmed fish, was transformed via electroporation with a plasmid expression vector driving the PhiX174 E-lysis gene under the transcriptional control by lambda PR regulatory sequence. The persistent maintenance of the plasmid vector in recombinant E. tarda was found in numerous subculture procedures over up to 6 months without any adverse effect on the original copy number of plasmids. Comparative examination based on semi-quantitative RT-PCR analysis on transcriptional efficiency of E-lysis gene between recombinant E. coli and E. tarda indicated that promoter strength and induction capacity of bacterial ghosts would be retarded in E. tarda as compared to the E. coli. However, the completeness of induction for bacterial ghosts in E. tarda was the same with E. coli, in which at least 99.99% of induction rate was possible and further the viability of recombinant bacteria was completely eliminated by a post-induction procedure including washing and freeze drying lyophilization.

Cloning and Nucleotide Sequence of the recA Gene from Shigella sonnei KNIH104S Isolated in Korea

  • Park, Yong-Chjun;Shin, Hee-Jung;Kim, Young-Chang
    • BMB Reports
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    • v.32 no.5
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    • pp.436-439
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    • 1999
  • Shigella sonnei is an important cause of human enteric infections. S. sonnei KNIH104S was previously reported to be isolated from Korean shigellosis patients. We cloned a 2.8-kb KpnI fragment containing the recA gene encoding a recombinase from the chromosomal DNA of S. sonnei KNIH104S. This recombinant plasmid was named pRAK28. E. coli HB101, a recA mutant, cannot grow on Luria-Bertani medium in the presence of the alkylating agent methylmethane sulfonate, however, E. coli HB101 harboring pRAK28 was found to grow on this medium. As far as we know, we are the first to sequence the recA gene from S. sonnei. This gene is composed of 1062 base pairs with an ATG initiation codon and a TAA termination codon. Nucleotide sequence comparison of the S. sonnei recA gene exhibited 99.7% and 99.5% identity with those of S. flexneri and E. coli, respectively.

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Studies on biological characters and plasmid profiles of Escherichia coli isolated from pigs (돼지 유래 대장균의 생물학적 특성과 plasmid profile에 대하여)

  • Jeong, Soo-kwan;Jeong, Suk-chan;Choi, Won-pil
    • Korean Journal of Veterinary Research
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    • v.30 no.3
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    • pp.287-295
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    • 1990
  • The purpose of this study was the examination for presence of pilus antigen, O serogroups, colicin production, antibiotic susceptibility and plasmid profiles among E coli isolated from diarrheal piglets and fattening pigs in Taegu province. Of 145 E coli isolated, 98 strains (67.4%) possesed pilus antigens which belonged to either K88 (47.6%), K99 (11.7%) or 987P (8.3%) types. Fifty-nine strains (40.7%) were classified into tenO serogroups and their types were O8 (22.0%), O20(16.9%), O141(15.3%), O9(10.2%), O45(10.2%), O139(8.5%), O064(6.8%), O149(5.0%), O157(3.4%), and O115(1.7%). Thirty-three strains (22.8%) were colicinogenic and 6 strains (4.1%) were hemolytic. One hundred and thirty-nine strains (95.9%) of 145 E coli isolates were resistant to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, tetracycline, rifampicin and nalidixic acid, alone or in combination thereof. Ninety strains (64.7%) of 139 drug resistant strains carried R factor (R) which were transferable to the recipient by conjugation. In gel electrophoresis for the isolation of plasmid DNA, the number of plasmid DNA band varied from 2 to 11 in 16 E coli with pilus antigen. It's molecular weight ranged from 1.0 to 60.0 megadalton.

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Identification of Microorganisms Isolated from Wound Regions of Chickens (계육의 창상부위에서 분리된 미생물의 동정)

  • Kim, Hyun-Jin;Chun, Ho-Hyun;Kim, Ju-Yeon;Jang, Sung-Ae;Lee, Bong-Duk;Chae, Hyun-Seok;Song, Kyung-Bin
    • Food Science and Preservation
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    • v.17 no.2
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    • pp.301-306
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    • 2010
  • We identified microorganisms causing skin disease in slaughtered chickens. Ten microbial strains were isolated from skin wound regions on the back and legs of chickens. Fatty acid composition analysis of the cell membranes of isolated bacteria identified five isolates of Shigella sonnei, Proteus mirabilis (2), and Escherichia coli (2). In addition, 16S rRNA sequencing indicated that S. sonnei (99%), P. mirabilis (99%), and E. coli (99%) were the strains responsible for skin wounds in chickens. Therefore, these three species may be the major pathogenic bacteria causing skin wounds on the back and legs of chickens.

Monoclonal Antibody Production against Piglet Diarrhea Agent (Enterotoxigenic E. coli) by Cell Fusion-Hybridoma Cell Technique (세포융합(細胞融合) 및 Hybridoma 세포작성(細胞作成)에 의한 항자돈백리(抗仔豚白痢) Monoclone항체(抗體)의 생산(生産))

  • Kim, Uh-ho;An, Soo-hwan;Yoon, Young-dhuk
    • Korean Journal of Veterinary Research
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    • v.27 no.2
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    • pp.259-267
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    • 1987
  • Enterotoxigenic E. coli (ETEC) cause an acute diarrhea (white scour) in both animals and humans. The disease process initially involves the adherence and colonization of the mucosal surface of the small intestine, followed by the elaboration of a heat-labile enterotoxin (LT) and/or heat-stable enterotoxin (ST). Intestinal adherence or colonization by ETEC is generally mediated by a specific surface-associated pilus (fimbrial) antigen that endows the bacteria with the capacity to adhere to epitherial cell surface. Fourteen monoclonal antibodies (MAbs) directed against pili antigens of ETEC were obtained by cell fusion/hybridoma technique. They were characterized by indirect immunofluorescence assay (IFA), and divided into four groups: specific to K99 antigen (group 1), cross-reactive with K99 and F41 antigens (group 2), specific to K88 antigen (group 3) and specific to 987P and K88 antigens (group 4), respectively. These MAbs demonstrated the distinct pili (K) antigens on the surface of ETEC by IFA, and could be utilized as diagnostic reagent for the identification of ETEC. When eighty-seven field isolates of E. coli from piglet with diarrhea were tested by group 3 MAb, fourty-two strains (48.3%) has K88 pilus antigen suggesting that this is one of the major pilus antigen of ETEC present in fifeld.

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A Molecular Biotechnology For Removal of Toxic Heavy Metals

  • Bang Sang-Weon;Clark Douglas S.;Keasling Jay D.
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.128-135
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    • 2000
  • The thiosulfate reductase gene (PhsABC) from Salmonella typhimurium was expressed in Escherichia coli in order to produce sulfide from inorganic thiosulfate and precipitate metals as metal sulfide complexes. A 5.1-kb DNA fragment containing the native phsABC and a 3.7-kb DNA fragment, excluding putative promoter and regulatory regions were inserted into expression vectors pTrc99A and pJB866, respectively. Upon expression of phsABC, E. coli DH5$\alpha$ harboring the phsABC constructs showed higher thiosulfate reductase activity and produced significantly more sulfide than the control strain (E. coli DH5$\alpha$) under both aerobic and anaerobic conditions. Among the four constructs, E. coli DH5$\alpha$ harboring pSB74 produced the highest level of thiosulfate reductase and removed most of heavy metals from solution under anaerobic conditions. In a mixture of 100 $\mu$M each of cadmium, lead, and zinc, the strain could remove $99\%$ of the total metals from solution within 10 hours. Cadmium was removed first, lead second, and zinc last. In contrast, a negative control did not produce any measurable sulfide and removed very little metals from solution. These results have important implications for removal of metals from wastewater contaminated with several metals.

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Asymmetric resolution of racemic styrene oxide using recombinant Escherichia coli harboring epoxide hydrolase of Rhodotorula glutinis (Rhodotorula glutinis 유래의 고효율 재조합 Epoxide Hydrolase를 이용한 라세믹 Styrene Oxide의 비대칭 광학분할)

  • Park, Kyu-Deok;Choi, Sung-Hee;Kim, Hee-Sook;Lee, Eun-Yeol
    • KSBB Journal
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    • v.23 no.5
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    • pp.369-374
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    • 2008
  • The effects of reaction temperature and the addition of various detergents on the enantioselective hyrolysis activity of the recombinant Escherichia coli containing the epoxide hydrolase (EH) gene of Rhodotorula glutinis were investigated for the production of enantiopure styrene oxide. The recombinant E. coli harboring the EH gene from R. glutinis exhibited the enantiopreference toward (R)-styrene oxide with the maximum hydrolytic activity of $165.04{\mu}mol/min/mg$ of dry cell weight (dcw). The addition of 0.5% (w/v) Tween 20 at $10^{\circ}C$ increased the initial hydrolysis rate and enantioselectivity by 1.45-fold and 2.0-fold, respectively. Enantiopure (S)-styrene oxide was prepared with 99% ee enantiopurity and 46.0% yield (theoretical yield=50%) from 20 mM racemic styrene oxide.

Sterilization of Escherichia coli Based on Nd: YAG Resonator with a Pulsed Xenon Flashlamp

  • Kim, Hee-Je;Kim, Dong-Jo;Hong, Ji-Tae;Xu, Guo-Cheng;Lee, Dong-Gil
    • Journal of Electrical Engineering and Technology
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    • v.6 no.2
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    • pp.275-279
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    • 2011
  • Sterilization of Escherichia coli (E. coli) is examined using a unique pulsed ultra-violet (UV) elliptical reactor based on Nd:YAG laser resonator, UV radiation from a pulsed xenon flashlamp. The light from the discharge has a broadband emission spectrum extending from the UV to the infrared region with a rich UV contained. Sterilization method by using the UV light is fast, environment-friendly and it does not cause secondary pollution. A Nd:YAG laser resonator having elliptical shape has advantage of concentrating the radiation of the UV light at two foci as the quart sleeve filled with E. coli. The primary objective of this research is to determine the important parameters such as pulse per second (pps), the applied voltage for sterilizing E. coli by using an UV elliptical reactor. From the experiment result, the sterilization effect of UV elliptical reactor is better than that of UV cylindrical reactor, and it can be 99.9% of sterilization at 800V regardless of the pps within 10 minutes.

D99 Type I Signal Peptidase Implicated Stabilizing the Protein Structure (Type I 신호펩디드 가수분해효소에 존재하는 D99 아미노산 잔기의 구조적 역할 가능성)

  • Sung, Meesook;Eunyoung Han;Lee, Hoyoung
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.140-144
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    • 2003
  • Type Ⅰ signal peptidase is an integral membrane protein that functions to cleave signal peptides from secreted and membrane proteins. The enzyme serves as a potential target for the development of novel antibacterial agents due to its unique physiological properties. Despite being one of the best characterized enzymes, the catalysis of Type Ⅰ signal peptidase still remains controversy over the catalytic serine/lysine dyad mechanism. It appears that the dyad proteases are generally less efficient than the prototypical serine/histidine/aspartic acid triad found in most enzymes, although Type Ⅰ signal peptidase is an exception to this rule. In this paper, we have proposed that Type Ⅰ signal peptidase may act as the serine/lysine/aspartic acid triad cataltytic mechanism. Therefore, the aspartic acid 99 residue in the E. coli signal peptidase was chosen and mutated to an alanine to see if there is any possible role of the aspartic acid in the catalytic function. Type Ⅰ signal peptidase D99A protein was inactive in vitro assay using the procoat synthesized by in vitro transcription translation. However, the mutant was active using a highly sensitive in vivo assay. Pulse-chase experiments show that the replacement of aspartic acid 99 with alanine results in a very unstable signal peptidase molecule. Therefore, we conclude that it is unlikely that the residue is directly involved in catalysis, but rather plays an important role in stabilizing the protein structure.