Crop & Food Research, Private Bag 4704, Christchurch, New Zealand.
Plant Cell Rep. 1996 Sep;15(12):958-62. doi: 10.1007/BF00231596.
Particle bombardment and Agrobacterium-mediated DNA delivery into immature embryos and microbulbs were used to investigate the expression of the uidA gene in in vitro onion cultures. Both methods were successful in delivering DNA and subsequent uidA expression was observed. Optimal transient β-glucuronidase activity was observed in immature embryos that had been pre-cultured for three days and bombarded at a distance of 3 cm from the stopping plate, under 25 in Hg vacuum, using 900-1300 psi rupture discs. The CaMV35S-uidA gene construct gave five fold higher transient β-glucuronidase activity than the uidA gene construct regulated by any of four other promoters initially chosen for high experession in monocotyledonous tissues.
利用粒子轰击和农杆菌介导的 DNA 导入法将 uidA 基因导入洋葱幼胚和小鳞茎,研究其在洋葱体外培养中的表达情况。两种方法都成功地将 DNA 导入洋葱幼胚和小鳞茎,并观察到 uidA 基因的表达。在预培养 3 天后,于 25 英寸汞柱真空下,使用 900-1300psi 的破裂盘,在距停止板 3cm 处对幼胚进行粒子轰击,可观察到最佳的瞬时β-葡萄糖醛酸酶活性。CaMV35S-uidA 基因构建体的瞬时β-葡萄糖醛酸酶活性比最初选择的四个可在单子叶组织中高效表达的启动子调控的 uidA 基因构建体高 5 倍。