( s. An example of this type of experimen...)Optimization of Biolistic Transformation Using the Helium-Driven PDS-1000/He System

s. An example of this type of experiment, in which the macrocarrier travel distance was varied along with the helium pressure and target distance, is shown in Table 4. Analysis of the three variables indicates that the maximum number of transformants is obtained with the sample at level 2 and the helium pressure between 900 and 1,500 psi. The macrocarrier travel distance had little effect on either increasing or decreasing the transformation efficiency. Similar results were found when the gap width was varied as a function of the helium pressure and target distance; neither increasing nor decreasing the gap width affected the number of transformants produced (results not shown). These results were further verified by changing either the gap width or the macrocarrier travel distance under conditions which were optimal for transformation (M17 tungsten microparticles, target at level 2, and helium pressure of 1,100 psi). Again, there was no significant effect on the number of yeast transformants (results not shown).

Discussion
Transient expression of the GUS gene in cauliflower stem sections and stable transformation of a yeast ura auxotroph with the wild type URA gene were used as model systems to identify those bombardment parameters which are important for optimization of Biolistic transformation. The parameters and their optimum conditions are summarized in Table 5 for both systems. In both cases, the most important parameter to optimize is target distance; there is a significant decrease in expression when the target level is changed from the optimum of 6 cm. This is apparent when cauliflower sections are transformed with M17 tungsten microparticles (Table 1) and when
'"/>

Source:

Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
Related biology technology :

1. Two-dimensional Optimization of a Semi-nested PCR for Detecting Listeria monocytogenes
2. PCR Optimization
3. PCR Optimization Kit
4. The Flexible matriXarray Chip System: A New Perspective for Probe Optimization in Gene-Expression Profiling
5. Rapid Chromatography Optimization by pH Scouting Using the New BioLogic Maximizer Valve System, Rev A
6. The Gradient Feature: Use in Optimization of Allelic Discrimination Assays, Rev B
7. Simple Approaches for Optimization of RT-PCR
8. Mobile Phase Optimization for the Analysis of an Antibody-Based Fusion Protein and Aggregates using a TSKgel Super SW3000 Size-Exclusion Column
9. Using the VersaFluor Fluorometer to Quantitate GUS Expression in Plant Tissues Bombarded With the Biolistic PDS-1000/He System
10. Biolistic Gene Transfer to Generate Transgenic Schistosomes, Rev A
11. Comparison of Performance Characteristics of Different Biolistic Devices