Epitopes fused to F-pilin are incorporated into functional recombinant pili

Journal of Molecular Biology
S RondotF Breitling

Abstract

In order to develop a system which allows infection by an epitope-specific phage-antibody via an F-pilus expressing that epitope, a study on the expression of foreign sequences on F-pilin was undertaken. Initially, a plasmid library was constructed with random sequences encoding one to five amino acid residues fused to the C terminus of F-pilin (traA) which was used to complement an F-plasmid with an amber mutation in traA. Functional F-pilin fusions were detected using the filamentous phage, fUSE2, which transduces tetracycline resistance, as well as immunoblots using a monoclonal antiserum specific for the acetylated N terminus of pilin. All the clones selected expressed the pilin-fusions and displayed full sensitivity towards fUSE2 infection, which was indistinguishable from the wild-type F-pilin. The sequences of fUSE2-sensitive clones when compared to randomly selected clones which were not fUSE2-sensitive, revealed no obvious pattern in the amino acid residues fused to the C terminus, except for a preference for a hydrophilic amino acid at position +1. Mutating the C-terminal Leu in wt (wild-type) pilin to Ser blocked pilus assembly and fUSE2 infection; the pilin was correctly processed but the level of acetylation at the...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon
Jul 27, 1990·Science·J K Scott, G P Smith
Dec 5, 1991·Journal of Molecular Biology·J D MarksG Winter
Sep 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·C F BarbasS J Benkovic
Aug 15, 1991·Gene·F BreitlingM Little
Jan 1, 1986·Annual Review of Genetics·K A Ippen-Ihler, E G Minkley
Jul 1, 1987·Journal of Bacteriology·K A SchandelR E Webster
Dec 1, 1985·Journal of Bacteriology·L S FrostJ S Lee
Dec 1, 1986·Microbiological Reviews·I Rasched, E Oberer
Dec 1, 1985·Molecular and Cellular Biology·G I EvanJ M Bishop
Jun 1, 1970·Journal of Bacteriology·J T Ou, T F Anderson
Jul 1, 1966·Proceedings of the National Academy of Sciences of the United States of America·L G Caro, M Schnös
Oct 1, 1984·Journal of Bacteriology·L S FrostN S Willetts
Jan 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·J McEwen, P Silverman
Jan 1, 1981·Molecular & General Genetics : MGG·D MooreK Ippen-Ihler
Sep 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·J D Boeke, P Model
Oct 15, 1983·Journal of Molecular Biology·M Chandler, D J Galas
Jan 1, 1980·Molecular & General Genetics : MGG·A KrämerE K Bautz
Nov 30, 1981·FEBS Letters·J ArmstrongJ E Walker
Jun 1, 1995·Journal of Bacteriology·K ManeewannakulK Ippen-Ihler
Nov 30, 1994·Annals of the New York Academy of Sciences·J A Francisco, G Georgiou
Jun 1, 1994·Microbiological Reviews·L S FrostR A Skurray
Mar 1, 1993·Journal of Bacteriology·K ManeewannakulK Ippen-Ihler
Apr 1, 1993·Journal of Bacteriology·D MaherD E Taylor
Mar 1, 1993·Journal of Bacteriology·D MooreK Ippen-Ihler
Feb 1, 1996·Molecular Immunology·M DueñasC A Borrebaeck
Jul 1, 1996·Journal of Bacteriology·N MajdalaniK Ippen-Ihler

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