Cloning of DNA sequences encoding foreign peptides and their expression in the K88 pili.
A genetic system that allows the cloning of a peptide-coding sequence in the Escherichia coli K88ac and K88ad pilin genes and their expression as recombinant pili has been constructed. Two insertion vectors were created by subcloning the pilin genes in a pBR322 plasmid and replacing the coding sequence of two nonconserved clusters by a linker. The K88ac helper genes were subcloned in the compatible pACYC184 plasmid, and expression of pili by bacteria carrying both plasmids occurred by complementation. Two peptide-coding sequences of the influenza hemagglutinin were cloned in both insertion vectors, and recombinant pilins were shown to be assembled in pili. One recombinant pilus was shown to elicit antibodies against the synthetic peptide in immunized rats. The somatostatin-coding sequence was cloned in both vectors and led in one case to detectable pilus production. The fused somatostatin was shown to be recognized by specific monoclonal and polyclonal antibodies.
Thiry
G
G
Central Laboratory, Solvay et Cie, Brussels, Belgium.
Clippe
A
A
Scarcez
T
T
Petre
J
J
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000235:genetics; D000954:Antigens, Surface / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics*; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology / Q000648:ultrastructure; D029968:Escherichia coli Proteins; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000648:ultrastructure*; D005786:Gene Expression Regulation; D005816:Genetic Complementation Test; D007118:Immunoassay; D015151:Immunoblotting; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D051381:Rats; D011919:Rats, Inbred Strains; D015183:Restriction Mapping
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D000939:Epitopes; D029968:Escherichia coli Proteins; C026888:K88 antigen, E coli; D010455:Peptides; D036781:Fimbriae Proteins