Rescue of a paralyzed-flagella mutant of Chlamydomonas by transformation

Proc Natl Acad Sci U S A. 1990 Aug;87(15):5739-43. doi: 10.1073/pnas.87.15.5739.

Abstract

The biflagellate alga Chlamydomonas has been used extensively in the genetic and biochemical analysis of flagellar assembly and motility. We have restored motility to a paralyzed-flagella mutant of Chlamydomonas by transforming with the corresponding wild-type gene. A nitrate reductase-deficient paralyzed-flagella strain, nit1-305 pf-14, carrying mutations in the genes for nitrate reductase and radial spoke protein 3, was transformed with wild-type copies of both genes. Two-thirds of the cells that survived nitrate selection also regained motility, indicating that they had been transformed with both the nitrate reductase and radial spoke protein 3 genes. Transformants typically contained multiple copies of both genes, genetically linked to each other, but not linked to the original mutant loci. Complementation of paralyzed-flagella mutants by transformation is a powerful tool for investigating flagellar assembly and function.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Blotting, Southern
  • Cell Movement
  • Chlamydomonas / genetics*
  • Chlamydomonas / physiology
  • DNA / genetics
  • DNA / isolation & purification
  • Electrophoresis, Gel, Two-Dimensional
  • Flagella / physiology
  • Flagella / ultrastructure
  • Mutation*
  • Phenotype
  • Plant Proteins / genetics
  • Plant Proteins / isolation & purification
  • Plasmids
  • Transformation, Genetic

Substances

  • Plant Proteins
  • DNA