Allelic subtyping of the intimin locus (eae) of pathogenic Escherichia coli by fluorescent RFLP.
Intimin is a highly polymorphic protein encoded by the eae gene and plays a crucial role in the attaching-effacing phenotype of diarrheagenic Escherichia coli and related pathogens. We have developed a method to quickly and accurately uncover allelic variation at the eae locus through the use of fluorescent RFLP (fRFLP). Application of fRFLP to 151 eae-positive strains (including the newly described Escherichia albertii) revealed 26 different fRFLP types that correspond to 20 of the 28 previously described eae alleles. Two sequence variants of the gamma, iota, kappa, and zeta alleles and three variants of epsilon were also observed. In addition to being reliable and accurate, the method can be easily adapted to accommodate new eae allelic sequences, as they become known.
Lacher
David W
DW
Microbial Evolution Laboratory, National Food Safety and Toxicology Center, Michigan State University, East Lansing, MI 48824, USA.
Steinsland
Hans
H
Whittam
Thomas S
TS
D018829:Adhesins, Bacterial / Q000145:classification / Q000235:genetics*; D000483:Alleles; D015373:Bacterial Typing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000145:classification / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000145:classification / Q000235:genetics*; D005456:Fluorescent Dyes; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
D018829:Adhesins, Bacterial; D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; C073819:eaeA protein, E coli