Isolation and Characterization of ARGONAUTE Mutants in Chlamydomonas

Methods Mol Biol. 2017;1640:159-172. doi: 10.1007/978-1-4939-7165-7_11.

Abstract

Random insertional mutagenesis and subsequent reverse genetic screening allow the isolation of mutants of interest. Here I describe the protocol for generating a tag insertion line and subsequent PCR-based screening for ARGONAUTE mutants as an example of a reverse genetic screen for the unicellular green alga Chlamydomonas reinhardtii.

Keywords: ARGONAUTE; Chlamydomonas; Genomic DNA extraction; Mutagenesis; PCR; miRNA.

MeSH terms

  • Argonaute Proteins / genetics*
  • Chlamydomonas reinhardtii / genetics*
  • DNA, Plant / genetics
  • Mutagenesis, Insertional / methods*
  • Mutation*
  • Plant Proteins / genetics*
  • Polymerase Chain Reaction / methods
  • Transformation, Genetic

Substances

  • Argonaute Proteins
  • DNA, Plant
  • Plant Proteins