Boston R S, Becwar M R, Ryan R D, Goldsbrough P B, Larkins B A, Hodges T K
Department of Botany and Plant Pathology, Purdue University, West Lafayette, Indiana 47907.
Plant Physiol. 1987 Apr;83(4):742-6. doi: 10.1104/pp.83.4.742.
Plasmids were constructed that contained promoters of ;plant' genes fused to the bacterial gene for chloramphenicol acetyl transferase. The promoters were isolated from a developmentally regulated Zea mays seed storage protein gene and from the mannopine synthase gene of the octopine type Ti plasmid of Agrobacterium tumefaciens which is constitutively expressed in crown gall tumors. These plasmids were introduced into carrot protoplasts made permeable by electroporation. Expression of chloramphenicol acetyl transferase activity directed by both promoters was positively correlated with DNA concentration. The efficiency of gene transfer was increased by inclusion of polyethylene glycol and by optimization of the voltage used in electroporation.
构建了质粒,这些质粒含有与氯霉素乙酰转移酶的细菌基因融合的“植物”基因启动子。这些启动子分别从一个受发育调控的玉米种子贮藏蛋白基因以及根癌土壤杆菌章鱼碱型Ti质粒的甘露碱合成酶基因中分离得到,后者在冠瘿瘤中组成型表达。通过电穿孔使胡萝卜原生质体具有通透性后,将这些质粒导入其中。两个启动子指导的氯霉素乙酰转移酶活性的表达与DNA浓度呈正相关。通过加入聚乙二醇以及优化电穿孔所用电压,提高了基因转移效率。