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Chloroplast Transformation and Reverse Genetics

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The Molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas

Part of the book series: Advances in Photosynthesis and Respiration ((AIPH,volume 7))

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Summary

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.

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Abbreviations

AAD:

aminoglycoside 3′′-adenyltransferase (or aminoglycoside 3′′-adenylyl transferase)

GUS:

β-Glucuronidase

PCR:

polymerase chain reaction

rDNA:

ribosomal RNA gene cluster

RFLP:

restriction fragment length polymorphism

rRNA:

ribosomal RNA

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Authors and Affiliations

  1. Departments of Molecular Biology and of Plant Biology, University of Geneva Sciences II, 30 quai E. Ansermet, 1211, Geneva 4, Switzerland

    Michel Goldschmidt-Clermont

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  1. Michel Goldschmidt-Clermont
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Editor information

Editors and Affiliations

  1. Departments of Molecular Biology and Plant Biology, University of Geneva, Geneva, Switzerland

    J. -D. Rochaix  & M. Goldschmidt-Clermont  & 

  2. Department of Chemistry and Biochemistry and Molecular Biology Institute, University of California-Los Angeles, Los Angeles, USA

    S. Merchant

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© 1998 Kluwer Academic Publishers

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Goldschmidt-Clermont, M. (1998). Chloroplast Transformation and Reverse Genetics. In: Rochaix, J.D., Goldschmidt-Clermont, M., Merchant, S. (eds) The Molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas. Advances in Photosynthesis and Respiration, vol 7. Springer, Dordrecht. https://doi.org/10.1007/0-306-48204-5_8

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  • DOI: https://doi.org/10.1007/0-306-48204-5_8

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-0-7923-5174-0

  • Online ISBN: 978-0-306-48204-5

  • eBook Packages: Springer Book Archive

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