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Mol Gen Genet. 2000 Apr;263(3):404-10.

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

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  • 1Department of Biology, University College London, UK.

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.

PMID:
10821174
[PubMed - indexed for MEDLINE]
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