【摘要】 根据最近文献报道的方法,以基因组测序用典型菌株Corynebacterium glutamicum ATCC13032为宿主,采用E.coli-C.glutamicum穿梭质粒pC2以及可以整合到谷氨酸棒杆菌基因组DNA上的质粒pAK进行电转化条件优化的实验,建立了一种简便的,适用于异源DNA高效整合转化的谷氨酸棒杆菌电转化方法。建立的新方法与原方法相比,细胞预培养时间缩短4h,细胞培养时间缩短1d左右。细胞培养基被简化,不再需要添加进口生化试剂。转化率可达5.5×106cfu／μgDNA,是一种适用于异源DNA高效整合转化的谷氨酸棒杆菌电转化法。更多还原

【Abstract】 Based on the recently reported method of electrotransformation of C . glutamicum , an improved method for integrative electrotransformation of C. glutamicum with Xenogeneic DNA was established with plasmids pC2 ( E. coli-C . glutamicum shuttle vector) and pAK(integrative vector for C . glutamicum) , and Corynebacterium glutamicum ATCC 13032 as the host cell. Two crucial alterations to the previously developed method are: (1) cultivation of cells for electrotransformation at 30 ℃ instead of 18 ℃; (2) no addition of isoniazid and imported reagent, Brain Heart Infusion, to media. The duration of cell precultivation was shortened by 4 hours and the time for cell cultivation cut down by about one day. A transformation efficiency of 5.5 × 106 cfu/μg DNA with Xenogeneic DNA was achieved with the method.更多还原