Sun L, Liang P, Tian Y, Mang K, Ba F, Ji P, Mei R
Institute of Microbiology, Chinese Academy of Sciences, Beijing.
Chin J Biotechnol. 1994;10(1):1-9.
Gram positive and negative bacterial shuttle vectors carrying Bacillus thuringiensis (B.t.) toxin gene were introduced by electroporation into wild-type Bacillus cereus, B. brevis and B. subtilis. The transformation efficiencies for these bacteria were around 10(1)-10(4) transformants per micrograms DNA based on the numbers of neomycin- and ampicillin-resistant colonies produced. The structure of the transferred plasmids proved identical with the original ones both in size and restriction pattern. Toxicity assays showed that the transformants gave a mortality of 90-100% against caterpillar of Heliothis assulta, indicating that the gene function was not changed by electroporation.
通过电穿孔法将携带苏云金芽孢杆菌(B.t.)毒素基因的革兰氏阳性和阴性细菌穿梭载体导入野生型蜡状芽孢杆菌、短短芽孢杆菌和枯草芽孢杆菌。根据产生的新霉素和氨苄青霉素抗性菌落数量,这些细菌的转化效率约为每微克DNA产生10(1)-10(4)个转化体。转移质粒的结构在大小和限制性图谱上均与原始质粒相同。毒性试验表明,转化体对烟青虫幼虫的死亡率为90-100%,这表明电穿孔法未改变基因功能。