TRANSIENT PLANT GENE-EXPRESSION - A SIMPLE AND REPRODUCIBLE METHOD BASED ON FLOWING PARTICLE GUN

被引:25
作者
GODON, C
CABOCHE, M
DANIELVEDELE, F
机构
[1] Laboratoire de Biologie Cellulaire, INRA, 78026 Versailles Cedex, route de Saint-Cyr
关键词
PARTICLE GUN; TRANSIENT EXPRESSION; 35S; BETA-GLUCURONIDASE; LUCIFERASE;
D O I
10.1016/0300-9084(93)90065-Z
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:591 / 595
页数:5
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