Godon C, Caboche M, Daniel-Vedele F
Laboratoire de Biologie Cellulaire, INRA, Versailles, France.
Biochimie. 1993;75(7):591-5. doi: 10.1016/0300-9084(93)90065-z.
Successful transient expression of beta-glucuronidase (Gus) and luciferase (Luc) in Nicotiana tabacum leaves was obtained after gene delivery by a simple and inexpensive particle gun. Takeuchi's biolistic process system was adapted to accelerate directly in a helium stream DNA-coated microprojectiles into intact plant leaf tissues. After bombardment of p70-omega-59Gus construction (duplication of the CaMV 35S enhancer), variability inherent to the bombardment procedure was observed in Gus activity measurements between replicate samples. We therefore included an internal standard (35S-Luc) in each bombardment, allowing an accurate, sensitive and reproducible comparison of the effective strength of different Gus-reporter constructs based on Gus/Luc ratio measurements.
通过一种简单且廉价的粒子枪进行基因传递后，在烟草叶片中成功实现了β-葡萄糖醛酸酶（Gus）和荧光素酶（Luc）的瞬时表达。竹内的生物弹道过程系统经过改良，可在氦气流中将包被了DNA的微弹直接加速打入完整的植物叶片组织。在用p70-omega-59Gus构建体（CaMV 35S增强子的重复序列）进行轰击后，在重复样本的Gus活性测量中观察到了轰击过程固有的变异性。因此，我们在每次轰击时都加入了一个内标（35S-Luc），从而能够基于Gus/Luc比率测量，对不同Gus报告基因构建体的有效强度进行准确、灵敏且可重复的比较。
