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

ase in the number of gold particles and in a proportional decrease in the number of transiently expressing cells; plates bombarded with 1 mg of 0.6 gold particles had more transiently-expressing anthocyanin cells than did plates bombarded with 1 mg of 1 particles. Tissue damage was less in cells bombarded with 0.6 gold than in tissue bombarded with the same amount of 1 gold. Although the presence of the wire mesh did not affect the number of cells that transiently express anthocyanin, it did reduce tissue damage and resulted in higher numbers of stable events. Results of stable transformation are presented in Table 2. Transformation was assessed by counting the number of individual multi-celled red-pigmented sectors present after 6 weeks of growth on non-selective media. Stable sectors are derived from cells which have been transformed, divide, and continue to express the anthocyanin gene in the daughter cells. Each stable sector has the potential of growing into a mature plant expressing the transforming gene. Compared to callus tissue bombarded with 1 gold, four- to eight-fold more red sectors were present in tissue bombarded with 0.6 gold, although in one experiment eight-fold less gold (and hence, eight-fold fewer gold particles) was used than in the other experiment (see Table 2). These results indicate that the level of transient expression is not always an accurate indication of the level of stable expression. Since the experiments summarized in Table 2 were carried out at different times and with different callus culture lines, the absolute numbers of transformants should not be compared between Experiment 1 and Experiment 2.

Stable transformation of Chlamydomonas. Chlamyd
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