Chimeric FimH adhesin of type 1 fimbriae: a bacterial surface display system for heterologous sequences.
The FimH adhesin of type 1 fimbriae has been tested as a display system for heterologous protein segments on the surface of Escherichia coli. This was carried out by introduction of restriction site handles (BglII sites) in two different positions in the fimH gene, followed by in-frame insertion of heterologous DNA segments encoding two reporter sequences. In the selected positions such insertions did not significantly alter the function of the FimH protein with regard to surface location and adhesive ability. The system seemed to be quite flexible, since chimeric versions of the FimH adhesin containing as many as 56 foreign amino acids were transported to the bacterial surface as components of the fimbrial organelles. Furthermore, the foreign protein segments were recognized by insert-specific antibodies when expressed within chimeric proteins on the surface of the bacteria. The results from this feasibility study point to the possibility of using the FimH adhesin as a general surface display system for sizeable protein segments.
Department of Microbiology, Technical University of Denmark, Lyngby, Denmark.
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Pallesen
L K
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Poulsen
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Christiansen
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P
Klemm
D018829:Adhesins, Bacterial / Q000235:genetics* / Q000378:metabolism; D018830:Adhesins, Escherichia coli; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D002772:Cholera Toxin / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D036781:Fimbriae Proteins; D015870:Gene Expression; D005798:Genes, Bacterial; D006514:Hepatitis B Surface Antigens / Q000235:genetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011498:Protein Precursors / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D018829:Adhesins, Bacterial; D018830:Adhesins, Escherichia coli; D017931:DNA Primers; D004269:DNA, Bacterial; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C058769:fimH protein, E coli; C050807:presurface protein 2, hepatitis B surface antigen; D036781:Fimbriae Proteins; D002772:Cholera Toxin; D004262:DNA Restriction Enzymes