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Construction of non-toxic Escherichia coli and Vibrio cholerae strains expressing high and immunogenic levels of enterotoxigenic E. coli colonization factor I fimbriae.

Artikel i vetenskaplig tidskrift
Författare Joshua Tobias
Michael Lebens
Ingrid Bölin
Gudrun Wiklund
Ann-Mari Svennerholm
Publicerad i Vaccine
Volym 26
Nummer/häfte 6
Sidor 743-52
ISSN 0264-410X
Publiceringsår 2008
Publicerad vid Institutionen för biomedicin, avdelningen för mikrobiologi och immunologi
Sidor 743-52
Språk en
Länkar dx.doi.org/10.1016/j.vaccine.2007.1...
Ämnesord Administration, Oral, Animals, Antibodies, Bacterial, blood, Bacterial Vaccines, administration & dosage, immunology, Enterotoxigenic Escherichia coli, immunology, Escherichia coli, immunology, metabolism, Escherichia coli Infections, blood, immunology, Female, Fimbriae Proteins, biosynthesis, genetics, Fimbriae, Bacterial, genetics, Formaldehyde, Immunization, Immunization Schedule, Immunoglobulin A, blood, Immunoglobulin G, blood, Immunoglobulin M, blood, Mice, Mice, Inbred BALB C, Operon, Plasmids, Protein Engineering, Recombinant Proteins, biosynthesis, Vaccines, Synthetic, administration & dosage, immunology, Vibrio cholerae, immunology, metabolism
Ämneskategorier Mikrobiologi inom det medicinska området

Sammanfattning

To express high quantities of colonization factor antigen I (CFA/I) derived from enterotoxigenic Escherichia coli (ETEC) for use in ETEC vaccines, the entire CFA/I operon consisting of four genes (cfa-A, -B, -C, -E) was cloned into plasmid expression vectors that could be maintained either with or without antibiotic selection. Expression from the powerful tac promoter was under the control of the lacIq repressor present on the plasmids. Fimbriae were expressed on the surface of both a non-toxigenic E. coli K12 strain and a non-toxigenic strain of Vibrio cholerae following induction with isopropyl-beta-D-thiogalactopyranoside (IPTG). It was found that the recombinant E. coli strains expressed up to 16-fold higher levels of CFA/I fimbriae compared to a reference strain which had previously been shown to be among the highest natural producers of the CFA/I fimbriae among tested wild type ETEC strains. Oral immunization with formalin-killed recombinant E. coli bacteria over-expressing CFA/I induced significantly higher serum IgA and IgG+M antibodies responses compared to the reference strain. Oral immunization with formalin-killed recombinant V. cholerae bacteria also induce strong CFA/I-specific serum IgA and IgG+M responses. We conclude that our constructs may be useful as candidate strains in an oral killed CF-ETEC vaccine.

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