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Optimization of Biolistic Transformation Using the Helium-Driven PDS-1000/He System

yeast are transformed with M10 and M17 tungsten or 1.0 mm gold microparticles (Table 3). This reduced transformation efficiency is most likely due to the lower velocity of the microparticles at longer travel distances.

Microparticle size also has a significant effect on transformation efficiency, probably for different reasons, however. In the case of cauliflower stem sections, when the optimum particle accelerator parameters were used, the 1.6 mm gold and M17 tungsten (median diameter 1.1 mm) microparticles produced more GUS-expressing cells than did the smaller 1.0 mm gold and M10 tungsten (median diameter 0.7 mm) microparticles. Furthermore, using the 1.6 mm gold microparticles, significant GUS expression was obtained when the cauliflower sections were bombarded at level 4. This may be attributable to the larger size of the 1.6 mm gold microparticles compared to the other microparticles assayed; the velocity of the larger microparticles is less affected by the travel distance. On the other hand, in the case of yeast transformation, using optimum particle accelerator parameters, the M17 tungsten microparticles were slightly better than the 1.0 mm gold and M10 tungsten microparticles in producing ura+ transformants. However, almost no transformants were found when the 1.6 mm gold microparticles were used. This effect is most likely the result of the larger 1.6 mm gold microparticles causing significant damage to the yeast cells (average diameter 6 mm) upon penetration. Because of the heterologous size distribution of the tungsten microparticles, their biological effect is due to a subset of small diameter microparticles.

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