Molecular characterization of a protective outer membrane lipoprotein (OmlA) from Actinobacillus pleuropneumoniae serotype 1
| Autor: | Amalia Rossi-Campos, C Anderson, Gerald-F. Gerlach, Andrew A. Potter, Sandy Klashinsky, Philip Willson |
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| Rok vydání: | 1993 |
| Předmět: |
Male
Swine Lipoproteins Molecular Sequence Data Immunology Molecular cloning medicine.disease_cause Microbiology law.invention Plasmid law medicine Animals Amino Acid Sequence Cloning Molecular Actinobacillus pleuropneumoniae Escherichia coli Peptide sequence Southern blot Base Sequence biology biology.organism_classification Molecular biology Recombinant Proteins Infectious Diseases Genes Bacterial Recombinant DNA Female Parasitology Rabbits Bacterial outer membrane Research Article Bacterial Outer Membrane Proteins |
| Zdroj: | Infection and Immunity. 61:565-572 |
| ISSN: | 1098-5522 0019-9567 |
| DOI: | 10.1128/iai.61.2.565-572.1993 |
| Popis: | An expression library was constructed from an Actinobacillus pleuropneumoniae serotype 1 clinical isolate using a plasmid vector. The library was screened with serum raised against the culture supernatant of this strain. One Escherichia coli transformant which also reacted with convalescent serum was isolated and found to express a protein with an electrophoretic mobility of approximately 50,000. The A. pleuropneumoniae-derived DNA encoding the protein was localized and characterized by nucleotide sequence analysis and primer extension mapping. One open reading frame of 1,095 bases was detected and confirmed by TnphoA insertion mutagenesis. It encoded a protein with a calculated molecular mass of 40 kDa which was lipid modified and present in the outer membrane and in membrane blebs of A. pleuropneumoniae. This protein was designated as outer membrane lipoprotein A (OmlA), and the encoding gene as omlA. Southern blotting under low-stringency conditions revealed the presence of hybridizing sequences in all A. pleuropneumoniae type strains, and a specific serum detected a homologous protein in serotypes 2, 8, 9, 11, and 12 type strains. Pigs immunized with this recombinant protein preparation were protected from death in an aerosol challenge experiment with an A. pleuropneumoniae serotype 1 isolate. |
| Databáze: | OpenAIRE |
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