Major improvements in gene delivery into mulberry leaf cells by particle inflow gun
2000
Sugimura, Y. (Kyoto Inst. of Technology (Japan)) | Uchida, A. | Adachi, T. | Hasegawa, H. | Kotani, E. | Furusawa, T.
Factors influencing the efficiency of gene delivery into mulberry leaf cells using a particle inflow gun were studied by employing a chimeric gene encoding for the production of beta-glucuronidase. Two days following bombardment with plasmid-coated microprojectiles, transient expression of the beta-glucuronidase was detected with the synthetic substrate (X-gluc), which, upon cleavage, formed a blue precipitate visually detectable within transformed cells. Bombardment efficiencies were determined by counting the number of blue spots that appeared in bombarded leaf tissue. The factors that remarkably increased bombardment efficiency were found to be the size of microprojectiles, the number of bombardments, the duration of preculture and the preconditioning of target tissue. When the optimized factors were combined, significantly high levels of bombardment efficiency were observed
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