Cloning and expression of two Pasteurella multocida genes in Escherichia coli.
A library of cloned Pasteurella multocida (toxigenic strain 9222, serotype D2) genomic sequences was constructed in Escherichia coli by incorporating TaqI digestion fragments into the plasmid vector pUC19. Immunological screening with antibodies directed against porin H, the major protein of the P multocida outer membrane, allowed the identification of a recombinant plasmid containing a 2.9-kbp DNA insert. This plasmid encoded the synthesis of two polypeptides, p25 (25 kDa) and p28 (28 kDa) which were detected in the different compartments of the E coli transformant. The peptide p25 was more abundant in the periplasm whereas p28 was mainly found in the cell envelope and in the cytosol. Immunological analysis indicates that p25, in contrast to p28, is antigenically related to porin H of P multocida. The expression in E coli of the gene encoding p28 was enhanced by induction of the lac promoter.
Département 'Membranes et Osmoregulation', CNRS-URA no 256, Université de Rennes I, France.
F
F
Manoha
G
G
Chevalier
H
H
Wróblewski
C
C
Delamarche
D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D015698:Genomic Library; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D016979:Pasteurella multocida / Q000235:genetics*; D010452:Peptide Biosynthesis; D010455:Peptides / Q000235:genetics*; D018272:Porins / Q000235:genetics* / Q000276:immunology; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D010455:Peptides; D018272:Porins