Isolation of Kluyveromyces lactis mannoprotein mutants by fluorescence-activated cell sorting.

Abstract

Wild type Kluyveromyces lactia cells bind 3 times more fluorescein-labeled wheat germ agglutinin (fl-WGA) than do mutants whose mannoprotein lacks terminal N-acetylglucosamine.This observation was used to develop a new procedure in which mannoprotein mutants are isolated by separating mutagenized cells labeled with 8-WGA using a fluorescence-activated cell sorter.When K. Iactis cells that bound tittle 8-WGA were selected by this procedure, mutants were obtained corresponding to the mnn2 class previously isolated by antibody agglutination methods (Smith, W. L., Naka-