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Sub-Micron Gold Particles Are Superior to Larger Particles for Efficient Biolistic Transformation of Organelles and Some Cells

Barbara Randolph-Anderson1, John E. Boynton1, John Dawson2, Erik Dunder2, Robert Eskes3, Nicholas W. Gillham1, Anita Johnson1, Philip S. Perlman3, Jan Suttie2, and William C. Heiser4.

1Developmental, Cell and Molecular Biology Group, Box 91000, Duke University, Durham, North Carolina 27708-1000; 2CIBAGEIGY Agricultural Biotechnology, P.O. Box 12257, Research Triangle Park, North Carolina 27709; 3Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas 75235-9038; 4Genetic Systems Division, Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules, California 94547


Summary
Stable transformation of maize, Chlamydomonas reinhardtii, and Saccharomyces cerevisiae nuclear genomes, as well as C. reinhardtii chloroplasts and S. cerevisiae mitochondria was achieved by particle bombardment with the PDS-1000/He instrument using plasmid DNA coated onto gold or tungsten microparticles. Results demonstrate that the kind and size of microparticles are important factors in determining the efficiency of transformation. Maize callus is transformed approximately five-fold more efficiently with 0.6 gold particles than with 1 gold particles. Nuclear transformation of S. cerevisiae is over ten-fold more efficient using 0.6 gold particles than with 1 gold particles. Yeast mitochondrial transformants, which arise at a very low frequency using 0.6 gold particles, were not found using larger gold particles or M5 tungsten particles. Nuclear transformation of Chlamydomonas was 2.5-fold more efficient with 0.6 gold particles than with either 1 gold or M10 tu
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