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Science 10 June 1988:
Vol. 240. no. 4858, pp. 1534 - 1538
DOI: 10.1126/science.2897716
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Articles

Science, Vol 240, Issue 4858, 1534-1538
Copyright © 1988 by American Association for the Advancement of Science

articles

Chloroplast transformation in Chlamydomonas with high velocity microprojectiles

JE Boynton, NW Gillham, EH Harris, JP Hosler, AM Johnson, AR Jones, BL Randolph-Anderson, D Robertson, TM Klein, KB Shark, and al. et

Department of Botany, Duke University, Durham, NC 27706.

Bombardment of three mutants of the chloroplast atpB gene of Chlamydomonas reinhardtii with high-velocity tungsten microprojectiles that were coated with cloned chloroplast DNA carrying the wild-type gene permanently restored the photosynthetic capacity of the algae. In most transformants of one of the mutants, a fragment with a 2.5-kilobase deletion was restored to normal size by a homologous replacement event; in about 25 percent of the transformants the restored restriction fragment was 50 to 100 base pairs smaller or larger than that of wild type. About one-fourth of the transformants of this mutant contained unintegrated donor plasmid when first examined. This plasmid persisted in four different transformants after 65 cell generations of continuous liquid culture but was lost from all transformants maintained on plates of selective medium. The restored wild-type atpB gene remains in all transformants as an integral part of the chloroplast genome and is expressed and inherited normally.


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