摘要
室温下用50mmol/LCaCl_2处理根癌土壤杆菌(Agrobacterium tumefaciens)以制备感受态细胞,然后经0℃冰浴及28℃热击处理,成功地将Ti质粒中间载体(>10kb)导入了根癌土壤农杆菌中。转化效率每个活细胞可达10^(-4)～10^(-5)转化子或10~6转化子/μgDNA。探讨了该菌细胞生长状态、CaCl_2滚滚浓度、温度、液氮、热击、复苏时间以及感受态细胞于4℃或—20℃(加15％甘油)下保存时间对根癌土壤杆菌转化的影响。
A successful transformation of Ti piasmid vector (>10kb) to Agrobaclerium tumefaciens (A. t) was described. Competent A. t cells were prepared with 50 mmol/L CaCl2 at room temperature and transformation was done in ice bath followed by 28℃ heat pulse. The transformation efficiency is 10-4-10-5 transiormants/recipient ceils or 106 transformsnts per ug DNA. Moreover, the effects of grown stage of A. t cells, concentration of CaCl2 solution,temperature,liquid N2,heat puise,recovery time and storage time of competent A. t cells at 4℃ or - 20℃ (mixed with 15% glycerin) on transformation were studied.
出处
《生物工程学报》
CAS
CSCD
北大核心
1995年第4期350-355,共6页
Chinese Journal of Biotechnology
