摘要
对去甲基化氮胞苷(5-AZA)的最佳浓度以及影响棉花角斑病菌[Xanthmonas campestris pv.malvacearum(Xcm)]V8电转化效率的诸多因素进行逐一研究,以期获得高效的电转化条件。结果表明,选取100μmol/L的5-AZA驯化处于对数中期的V8制备感受态细胞;保证电压2.1 kV、50μg/mL质粒7μL、4℃预冷复苏培养基SOC、振荡复苏培养4~5 h时,V8的电转化效率高达5×10^2/μg DNA。
In order to obtain efficient electrical conversion conditions,experimentswere conducted to study the efficiency between optimal concentration for 5-AZA and factors playing roles in the efficiency of cotton keratinae[Xanthmonas campestris pv.malvacearum(Xcm)]V8 one by one.The results showed that V8 for preparing competent cells was domestricated by 5-AZA with a concentration of 100μmol/L at the logarithmic metaphase,Whiah was used to acclimate V8 in the medium logarithm to prepare competent cells.When the guaranteed voltage was 2.1 kV,7μL plasmid as well as concentration of 50μg/mL,recovery media SOC of 4℃ pre-cooling,culturing with time of four to five hours by oscillating,the efficiency could reach about 5×10^2/μg DNA.
作者
王清
孟凡奇
张勇跃
缪卫国
刘志坚
WANG Qing;MENG Fan-qi;ZHANG Yong-yue;MIAO Wei-guo;LIU Zhi-jian(Luohe Academy of Agricultural Sciences,Luohe 462300,Henan,China;Key Laboratory of Green Preventionand Control ofTropical Plant Diseases and Pests(Hainan University),Ministry of Education,Haikou 570228,China)
出处
《湖北农业科学》
2020年第12期175-178,共4页
Hubei Agricultural Sciences
基金
国家甘薯产业技术体系项目(CARS-10-C-13)
国家重大基础研究计划项目(2011CB111612)
国家农业产业技术体系建设项目(CARS-34-GW8)。
