title: Knocking out the wall: protocols for gene targeting in Physcomitrella patens.
abstract: The moss Physcomitrella patens has become established as a model for investigating plant gene function due to the feasibility of gene targeting. The chemical composition of the P. patens cell wall is similar to that of vascular plants and phylogenetic analyses of glycosyltransferase sequences from the P. patens genome have identified genes that putatively encode cell wall biosynthetic enzymes, providing a basis for investigating the evolution of cell wall polysaccharides and the enzymes that synthesize them. The protocols described in this chapter provide methods for targeted gene knockout in P. patens, from constructing vectors and maintaining cultures to transforming protoplasts and analyzing the genotypes and phenotypes of the resulting transformed lines.
authors.affiliation: Department of Biological Sciences, Center for Biotechnology and Life Sciences, University of Rhode Island, Kingston, RI, USA.
authors.forename: Alison W
authors.initials: AW
authors.lastname: Roberts
authors.forename: Christos S
authors.initials: CS
authors.lastname: Dimos
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Budziszek
authors.forename: Chessa A
authors.initials: CA
authors.lastname: Goss
authors.forename: Virginia
authors.initials: V
authors.lastname: Lai
mesh_terms: D019068:Bryopsida / Q000166:cytology / Q000235:genetics*; D018929:Cell Culture Techniques; D002473:Cell Wall / Q000737:chemistry / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D055786:Gene Knockout Techniques; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005838:Genotype; D030821:Plants, Genetically Modified / Q000166:cytology / Q000235:genetics; D011523:Protoplasts