Cloning and characterization of the gene coding for NADPH-sulfite reductase hemoprotein from Actinobacillus pleuropneumoniae and use of the protein product as a vaccine
Pj. Willson et al., Cloning and characterization of the gene coding for NADPH-sulfite reductase hemoprotein from Actinobacillus pleuropneumoniae and use of the protein product as a vaccine, CAN J VET R, 65(4), 2001, pp. 206-212
Citations number
30
Categorie Soggetti
Veterinary Medicine/Animal Health
Journal title
CANADIAN JOURNAL OF VETERINARY RESEARCH-REVUE CANADIENNE DE RECHERCHE VETERINAIRE
An expression library was constructed from an Actinobacillus pleuropneumoni
ae serotype 1 clinical isolate and screened with serum produced in pigs tha
t had been vaccinated with the anionic fraction of a sodium chloride extrac
t. One E. coli transformant was isolated that produced a large amount of a
protein with an electrophoretic mobility of about 67 000 molecular mass. Th
e A. pleuropneumoniae-derived DNA encoding the protein was localized and ch
aracterized by restriction enzyme digestion and nucleotide sequence analysi
s which showed strong homology with the cysI gene of E. coli. One open read
ing frame of 1764 bases in length was detected which encoded a cysI protein
from serotype 1, with a calculated molecular mass of 66 678. The DNA encod
ing the protein was labeled with radio-isotope and the homologous gene was
isolated from an A. pleuropneumoniae serotype 5a library. The serotype 5a g
ene was the same length, but the cysI protein from serotype 5a was slightly
larger (66 849) due to 8 substitutions in the amino acid sequence. Express
ion plasmids containing cysI from either serotype of A. pleuropneumoniae co
mplemented an E. coli cysI mutant. Pigs vaccinated with the recombinant cys
I were protected from challenge with A. pleuropneumoniae of the homologous
serotype.