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OALib Journal期刊
ISSN: 2333-9721
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恶臭假单胞菌kt2440高效电转化方法
DOI: 10.3724/SP.J.1145.2010.00432, PP. 432-434
Keywords: 恶臭假单胞菌kt2440,电转化,感受态细胞
Abstract:
恶臭假单胞菌（pseudomonasputida）kt2440是模式环境微生物菌株，也是异源表达的良好宿主菌.高效率的电转化方法对于基因的引入及后续实验非常关键.通过条件优化，建立了高效的电转化方法.em培养基培养菌体至d600nm=0.6~0.75，冰冷的3mmol/l4-羟乙基哌嗪乙磺酸（hepes，ph=7.0）的缓冲液洗涤菌体3次，菌体浓缩300倍，50μl分装并用于一次电转化.电转化条件：1.2kv，200ω，25μf.1mlsoc培养基悬浮后，转至15ml聚丙烯离心管培养2h,转化效率可高达3.0×108转化子/μg，比目前为止文献报道的该菌株电转化效率高出30倍左右.实验结果还表明，转化dna浓度与转化子数目呈线性关系.图2表1参14
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