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KMID : 0903619980390020222
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Journal of the Korean Society for Horticultural Science
1998 Volume.39 No. 2 p.222 ~ p.226
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Factors Affecting Transient Expression on Leaves of Phlox paniculate L. by Using Biolistic Gun
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Abstract
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Factors related to higher frequency of transient expression via helium driven Biolistic PDS/He 1000 system have been examined by using leaves of phlox (Phlox Paniculata L.). The optimal pressures for accelerating particles were 1,550 and 1,800 psi for increasing transient GUS (β-glucuronidase) gene expression. CaCl₂ and spermidine during preparation were required for bindig DNA on gold particles. The combination of longer vortexing time from 2 to 20 min and removal of solution using a piper instead of centrifugation for collecting coated particles reduced or eliminated the agglomerates, and thus, enhanced the frequency of GUS gene expression. By repetition based on modified protocols, GUS expression was detected on 44% of the total bombarded leaves and 3 blue foci per leaves were observed.
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