Molecular and General Genetics MGG

, Volume 263, Issue 3, pp 404–410

Tools for chloroplast transformation in Chlamydomonas: expression vectors and a new dominant selectable marker

  • J. M. Bateman
  • S. Purton
ORIGINAL PAPER

DOI: 10.1007/s004380051184

Cite this article as:
Bateman, J. & Purton, S. Mol Gen Genet (2000) 263: 404. doi:10.1007/s004380051184

Abstract

Reverse-genetic studies of chloroplast genes in the green alga Chlamydomonas reinhardtii have been hampered by the paucity of suitable selectable markers for chloroplast transformation. We have constructed a series of vectors for the targeted insertion and expression of foreign genes in the Chlamydomonas chloroplast genome. Using these vectors we have developed a novel selectable marker based on the bacterial gene aphA-6, which encodes an aminoglycoside phosphotransferase. The aphA-6 marker allows direct selection for transformants on medium containing either kanamycin or amikacin. The marker can be used to inactivate or modify specific chloroplast genes, and can be used as a reporter of gene expression. The availability of this marker now makes possible the serial transformation of the chloroplast genome of Chlamydomonas.

Key wordsChlamydomonasChloroplast transformationaphA-6Selectable marker

Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  • J. M. Bateman
    • 1
  • S. Purton
    • 1
  1. 1.Department of Biology, University College London, Gower Street, London, WC1E 6BT, UK Tel.: +44-171-5042675; Fax: +44-171-380709 E-mail: s.purton@ucl.ac.ukGB