( iolistic transformation is t...)Optimization of Biolistic Transformation Using the Helium-Driven PDS-1000/He System

iolistic transformation is the helium pressure. While the fungal and plant experiments reported here show that the range over which optimum transformation occurs is quite broad (there is little difference in transformation efficiency between 900 and 1,550 psi), this is not always the case. In other systems, such as the stable transformation of bacteria (Smith, et al., 1992) or plants (Russell, et al., 1992a), the optimum helium pressure range is narrower. Because microparticle bombardment will result in cell damage, the most gentle conditions which give a high transformation efficiency should be used. This is particularly true when transient gene assays are performed as preliminary experiments for determining conditions to use in generating stable transformants (see below).

The experimental results obtained here indicate that two of the parameters which can be varied on the PDS-1000/He instrument, the gap distance and the macrocarrier travel distance, may have little effect on the transformation efficiency. Varying either of these parameters and subsequently assaying their effect on yeast transformation (Table 2) or transient expression in cauliflower (Table 4) failed to show a significant effect. Table 6 summarizes the effect of changing the particle accelerator parameters on microparticle velocity and transformation efficiency.

Biolistic transformation of yeast and cauliflower stem sections has not been reported previously using the PDS-1000/He instrument. However, transformation of yeast and other plant tissues has been reported using the gunpowder-driven PDS- 1000 unit. Armaleo, et al. (1990) described optimized biological parameters for yeast, and recommended using 1.1 mm tungsten micropa
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