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      KCI등재 SCIE SCOPUS

      Protective Immune Response of Bacterially-Derived Recombinant FaeG in Piglets

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      https://www.riss.kr/link?id=A104674099

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      다국어 초록 (Multilingual Abstract)

      FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli(ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, C83907 (K88ad, CT+, ST+). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.
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      FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli(ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine ...

      FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli(ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, C83907 (K88ad, CT+, ST+). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

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      참고문헌 (Reference)

      1 Studier, "Use of T7 RNA polymerase to direct expression of cloned genes Methods in Enzymology" Academic Press 60-89, 1990

      2 Bakker, "Structure and function of periplasmic chaperone-like proteins involved in the biosynthesis of K88 and K99 fimbriae in enterotoxigenic Escherichia coli" 875-886, 1991

      3 Van den Broeck, W, "Receptor-dependent immune responses in pigs after oral immunization with F4 fimbriae" 67 : 520-526, 1999

      4 Ubeidat, M, "Purification and renaturation of Dictyostelium recombinant alkaline phospha-tase by continuous elution electrophoresis" 27 : 375-383, 2003

      5 Rutter, "Protection against enteric disease caused by Escherichia coli a model for vaccination with a virulence determinant? Nature 242" 531-532, 1973

      6 Furer, "Protection against colibacillosis in neonatal piglets by immunization of dams with procholeragenoid" 887-894, 1982

      7 Huang, Y.H, "Production of FaeG the major subunit of K88 fimbriae in transgenic tobacco plants and its immunogenicity in mice" 71 : 5436-5439, 2003

      8 Barman, N.N, "Passive immunization of piglets against enterotoxigenic colibacillosis by vaccinating dams with K88ac pili bearing bacterins" 37 : 1132-1135, 1999

      9 and A, "Monoclonal antibodies against the different subcomponents of colonization factor antigen II of enterotoxigenic Escherichia coli" 19901906-1912

      10 Mol, O, "Molecular and structural aspects of fimbriae biosynthesis and assembly in Escherichia coli" 19 : 25-52, 1996

      1 Studier, "Use of T7 RNA polymerase to direct expression of cloned genes Methods in Enzymology" Academic Press 60-89, 1990

      2 Bakker, "Structure and function of periplasmic chaperone-like proteins involved in the biosynthesis of K88 and K99 fimbriae in enterotoxigenic Escherichia coli" 875-886, 1991

      3 Van den Broeck, W, "Receptor-dependent immune responses in pigs after oral immunization with F4 fimbriae" 67 : 520-526, 1999

      4 Ubeidat, M, "Purification and renaturation of Dictyostelium recombinant alkaline phospha-tase by continuous elution electrophoresis" 27 : 375-383, 2003

      5 Rutter, "Protection against enteric disease caused by Escherichia coli a model for vaccination with a virulence determinant? Nature 242" 531-532, 1973

      6 Furer, "Protection against colibacillosis in neonatal piglets by immunization of dams with procholeragenoid" 887-894, 1982

      7 Huang, Y.H, "Production of FaeG the major subunit of K88 fimbriae in transgenic tobacco plants and its immunogenicity in mice" 71 : 5436-5439, 2003

      8 Barman, N.N, "Passive immunization of piglets against enterotoxigenic colibacillosis by vaccinating dams with K88ac pili bearing bacterins" 37 : 1132-1135, 1999

      9 and A, "Monoclonal antibodies against the different subcomponents of colonization factor antigen II of enterotoxigenic Escherichia coli" 19901906-1912

      10 Mol, O, "Molecular and structural aspects of fimbriae biosynthesis and assembly in Escherichia coli" 19 : 25-52, 1996

      11 Sambrook, J, "Molecular Cloning: A Laboratory Manual3rded." Cold Spring Harbor Laboratory Press 2001

      12 Van den Broeck, W, "Induction of immune responses in pigs following oral administration of purified F4 fimbriae" 17 : 2020-2029, 1999

      13 A, "Identification and characterization of precursors in the biosynthesis of the K88ab fimbria of Escherichia coli" 41-49, 1983

      14 Levine, "Escherichia coli that cause diarrhea" 377-389, 1987

      15 Greenwood, "Development and protective efficacy of a recombinant-DNA derived fimbrial vaccine against enterotoxic colibacillosis in neonatal piglets" 389-6 392, 1988

      16 Turnes, C.G, "DNA inoculation with a plasmid vector carrying the faeG adhesin gene of Escherichia coli K88ab induced imune responses in mice and pigs" 17 : 2089-2095, 1999

      17 Wouters, "Construction and characterization of mutants impaired in the biosynthesis of the K88ab antigen" 512-521, 1982

      18 Huang, Y.H, "Cloning and expression of the K99 fimbrial antigen gene of Enterotoxingenic Escherichia coli" 21 : 471-474, 2001

      19 Hong, "Cloning and expression of genetic determinants for K88 antigen" 36-45, 1985

      20 Willemsen, "Age and serotype dependent binding of K88 fimbriae to porcine intestinal receptors" 367-375, 1992

      21 Wizemann, T.M, "Adhesins as targets for vaccine development." 5 : 395-403, 1999

      22 Girardeau, "A new in vitro technique for attachment to intestinal villi using enteropathogenic Escherichia coli" 31-37, 1980

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      기준연도 WOS-KCI 통합IF(2년) KCIF(2년) KCIF(3년)
      2016 1.76 0.2 1.22
      KCIF(4년) KCIF(5년) 중심성지수(3년) 즉시성지수
      0.91 0.73 0.399 0.07
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