Section of Plant Biology, Division of Biological Sciences, Cornell University, Ithaca, NY 14853.
Proc Natl Acad Sci U S A. 1986 Sep;83(18):6815-9. doi: 10.1073/pnas.83.18.6815.
The bacterial chloramphenicol acetyltransferase (CAT) gene was expressed in protoplasts of three important graminaceous plant species after introduction of the gene by electroporation. Gene transfer occurred when high-voltage electric pulses were applied either directly or indirectly (without anode contact) to a solution containing plasmid DNA and protoplasts of rice, wheat, or sorghum. The indirect method was more rapid, resulted in higher protoplast viability, and was less subject to contamination than the direct-contact method. Gene expression of approximately equal magnitude resulted when the CAT gene was fused to either the 35S promoter of cauliflower mosaic virus or the copia long terminal repeat promoter of Drosophila. Together with recent advances in regeneration of callus and whole plants from protoplasts, this system makes it possible to study inheritance and expression of genes introduced into graminaceous monocotyledonous plants.
电穿孔法将细菌氯霉素乙酰转移酶(CAT)基因导入原生质体后,该基因在三种重要的禾本科植物中得到表达。当高电压电脉冲直接或间接(无阳极接触)施加到含有质粒 DNA 和水稻、小麦或高粱原生质体的溶液中时,基因转移就会发生。间接法比直接接触法更快,原生质体活力更高,污染程度也更低。当 CAT 基因与花椰菜花叶病毒 35S 启动子或果蝇 copia 长末端重复启动子融合时,CAT 基因的表达量大致相同。与原生质体愈伤组织和整株植物再生的最新进展相结合,该系统使得研究导入禾本科单子叶植物的基因的遗传和表达成为可能。