title: F4-related mutation and expression analysis of the aminopeptidase N gene in pigs.
abstract: Intestinal infections with F4 enterotoxigenic Escherichia coli (ETEC) are worldwide an important cause of diarrhea in neonatal and recently weaned pigs. Adherence of F4 ETEC to the small intestine by binding to specific receptors is mediated by F4 fimbriae. Porcine aminopeptidase N (ANPEP) was recently identified as a new F4 receptor. In this study, 7 coding mutations and 1 mutation in the 3' untranslated region (3' UTR)were identified in ANPEP by reverse transcriptase (RT-) PCR and sequencing using 3 F4 receptor-positive (F4R+) and 2 F4 receptor-negative (F4R-) pigs, which were F4 phenotyped based on the MUC4 TaqMan, oral immunization, and the in vitro villous adhesion assay. Three potential differential mutations (g.2615C > T, g.8214A > G, and g.16875C > G) identified by comparative analysis between the 3 F4R+ and 2 F4R- pigs were genotyped in 41 additional F4 phenotyped pigs. However, none of these 3 mutations could be associated with F4 ETEC susceptibility. In addition, the RT-PCR experiments did not reveal any differential expression or alternative splicing in the small intestine of F4R+ and F4R- pigs. In conclusion, we hypothesize that the difference in F4 binding to ANPEP is due to modifications in its carbohydrate moieties.
authors.affiliation: Laboratory of Animal Genetics, Faculty of Veterinary Medicine, Ghent University, Heidestraat 19, B-9820 Merelbeke, Belgium.
authors.forename: T
authors.initials: T
authors.lastname: Goetstouwers
authors.forename: M
authors.initials: M
authors.lastname: Van Poucke
authors.forename: V U
authors.initials: VU
authors.lastname: Nguyen
authors.forename: V
authors.initials: V
authors.lastname: Melkebeek
authors.forename: A
authors.initials: A
authors.lastname: Coddens
authors.forename: D
authors.initials: D
authors.lastname: Deforce
authors.forename: E
authors.initials: E
authors.lastname: Cox
authors.forename: L J
authors.initials: LJ
authors.lastname: Peelman
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D018826:CD13 Antigens / Q000235:genetics / Q000378:metabolism*; D054307:Enterotoxigenic Escherichia coli / Q000145:classification* / Q000502:physiology*; D004927:Escherichia coli Infections / Q000235:genetics / Q000382:microbiology / Q000662:veterinary*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D020022:Genetic Predisposition to Disease; D005838:Genotype; D009154:Mutation; D013552:Swine / Q000235:genetics* / Q000378:metabolism
chemical_list: D018826:CD13 Antigens