State key Lab of Microbial Technology, Shandong University, Jinan 250100, China.
J Microbiol Biotechnol. 2010 Jan;20(1):39-44.
Acidithiobacillus caldus is an acidophilic, chemoautotrophic bacterium, plays important role in bioleaching. Gene transformation into A. caldus is difficult and only the conjugation method was reported successful, which method was relatively sophisticated. In this research, electrotransformation of A. caldus species was achieved for the first time using A. caldus Y-3 and plasmid pJRD215. Transformants were confirmed by colony PCR specific to the str gene on pJRD215, and the recovery of the plasmid from the presumptive transformants. Optimizations were made and the transformation efficiency was increased from 0.8 to 3.6 x 104 transfomants/microng plasmid DNA. The developed electrotransformation method was convenient in introducing foreign genes into A. caldus.
嗜酸氧化亚铁硫杆菌是一种嗜酸、化能自养细菌，在生物浸出中起着重要作用。将基因转入嗜酸氧化亚铁硫杆菌比较困难，目前仅报道了接合方法成功，该方法相对复杂。本研究首次用电穿孔法实现了嗜酸氧化亚铁硫杆菌 Y-3 种和质粒 pJRD215 的转化。通过对 pJRD215 上 str 基因的菌落 PCR 特异性，以及从假定转化体中回收质粒，证实了转化体的存在。进行了优化，使转化效率从每微克质粒 DNA 0.8×104 个转化体提高到 3.6×104 个转化体。所建立的电穿孔转化方法为将外源基因导入嗜酸氧化亚铁硫杆菌提供了便利。
