title: Reverse genetics in Chlamydomonas: a platform for isolating insertional mutants.
abstract: A method was developed to identify insertional mutants of Chlamydomonas reinhardtii disrupted for selected target genes. The approach relies on the generation of thousands of transformants followed by PCR-based screenings that allow for identification of strains harboring the introduced marker gene within specific genes of interest. Our results highlight the strengths and limitations of two independent screens that differed in the nature of the marker DNA used (PCR-amplified fragment containing the plasmid-free marker versus entire linearized plasmid with the marker) and in the strategies used to maintain and store transformants.
authors.affiliation: Department of Plant Biology, The Carnegie Institution for Science, Stanford, CA 94305, USA. q62gobad@uco.es.
authors.forename: David
authors.initials: D
authors.lastname: Gonzalez-Ballester
authors.forename: Wirulda
authors.initials: W
authors.lastname: Pootakham
authors.forename: Florence
authors.initials: F
authors.lastname: Mus
authors.forename: Wenqiang
authors.initials: W
authors.lastname: Yang
authors.forename: Claudia
authors.initials: C
authors.lastname: Catalanotti
authors.forename: Leonardo
authors.initials: L
authors.lastname: Magneschi
authors.forename: Amaury
authors.initials: A
authors.lastname: de Montaigu
authors.forename: Jose J
authors.initials: JJ
authors.lastname: Higuera
authors.forename: Matthew
authors.initials: M
authors.lastname: Prior
authors.forename: Aurora
authors.initials: A
authors.lastname: Galván
authors.forename: Emilio
authors.initials: E
authors.lastname: Fernandez
authors.forename: Arthur R
authors.initials: AR
authors.lastname: Grossman