title: Use of micromanipulation and "feeder layers" to clone the oyster pathogen Perkinsus marinus.
abstract: Genetic and biochemical characterization of microbes often requires the use of clonal cultures. A method to clone the oyster parasite Perkinsus marinus is described. Individual cells are isolated via micromanipulation and maintained above an actively proliferating "feeder layer" of P. marinus on a 0.45-microm membrane. Extracellular products released from the proliferating feeder layer can diffuse across the membrane and bathe the isolated cell, stimulating it to proliferate. The method is relatively simple and should be applicable to most protists that can be cultured in the laboratory.
authors.affiliation: Baruch Marine Field Laboratory, Baruch Institute for Marine Biology and Coastal Research, University of South Carolina, Georgetown 29442 USA. bushek@sc.edu
authors.forename: D
authors.initials: D
authors.lastname: Bushek
authors.forename: R A
authors.initials: RA
authors.lastname: Holley
authors.forename: K S
authors.initials: KS
authors.lastname: Reece
mesh_terms: D000818:Animals; D016782:Apicomplexa / Q000254:growth & development* / Q000302:isolation & purification; D003470:Culture Media; D010125:Ostreidae / Q000469:parasitology*
chemical_list: D003470:Culture Media