Title

Forward and reverse genetic analysis of microtubule motors in Chlamydomonas

UMMS Affiliation

Department of Cell Biology; Program in Molecular Medicine

Date

1-3-2001

Document Type

Article

Subjects

Animals; Chlamydomonas; Dynein ATPase; Microtubules; Molecular Motor Proteins; Mutagenesis, Insertional; Mutation

Abstract

The ability to integrate biochemical, cell biological, and genetic approaches makes Chlamydomonas reinhardtii the premier model organism for studies of the eukaryotic flagellum and its associated molecular motors. Hundreds of motility mutations have been identified in Chlamydomonas, including many that affect dyneins and kinesins. These mutations have yielded much information on the structure and function of the motors as well as the roles of individual subunits within the motors. The development of insertional mutagenesis has opened the door to powerful new approaches for genetic analysis in Chlamydomonas. Insertional mutants are created by transforming cells with DNA-containing selectable markers. The DNA is randomly integrated throughout the genome and usually deletes part of the chromosome at the site of insertion, thereby creating mutations that are marked by the integrated DNA. These mutations can be used for forward genetic approaches where one characterizes a mutant phenotype and then clones the relevant gene using the integrated DNA as a tag. The insertional mutants also may be used in a reverse genetic approach in which mutants lacking a gene of interest are identified by DNA hybridization. We describe methods to generate and characterize insertional mutants, using mutations that affect the outer dynein arm as examples.

Rights and Permissions

Citation: Methods. 2000 Dec;22(4):285-98. Link to article on publisher's site

Related Resources

Link to Article in PubMed

Journal Title

Methods (San Diego, Calif.)

PubMed ID

11133235