Book chapter

Chloroplast transformation and reverse genetics

Published inThe molecular biology of chloroplasts and mitochondria in Chlamydomonas, Editors Rochaix, Jean-David ; Goldschmidt-Clermont, Michel & Merchant, S., p. 139-149
PublisherDordrecht : Kluwer
  • Advances in photosynthesis; 7
Publication date1998

For chloroplast transformation, the most efficient method to introduce DNA is particle bombardment. To then select the cells which harbor a transformed plastid, two classes of markers are available. With one class, selection is based on the rescue of a non-photosynthetic mutant with the wild-type chloroplast gene. With the other class, selection is based on a mutation or a foreign gene conferring resistance to an antibiotic or a herbicide. Transforming DNA is integrated by homologous recombination, and only in exceptional cases is it maintained extrachromosomally. The modified and wild-type copies of the highly polyploid plastid genome usually segregate rapidly, although in some circumstances a heteroplasmic mixture of genomes is retained. The available technology and markers readily allow chloroplast gene inactivation and site-directed mutagenesis. These possibilities are enhanced by strategies such as co-transformation or the repeated use of unstable markers.

Citation (ISO format)
GOLDSCHMIDT-CLERMONT, Michel P. Chloroplast transformation and reverse genetics. In: The molecular biology of chloroplasts and mitochondria in Chlamydomonas. Dordrecht : Kluwer, 1998. p. 139–149. (Advances in photosynthesis)
Main files (1)
Book chapter (Published version)
  • PID : unige:173855

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