摘要
采用高压电穿孔法将穿梭质粒导入了嗜热脂肪芽胞杆菌（Ｂａｃｉｌｌｕｓ ｓｔｅａｒｏｔｈｅｒｍｏｐｈｉｌｕｓ）Ｋ１０４１和Ｔ５２１菌株。以对数生长后期的菌体制备Ｋ１０４１转化细胞，以ＬＢ平板上于５０℃培养的过夜菌制备Ｔ５２１转化细胞，细胞密度为５～７×１０９细胞／ｍＬ。电击条件如下：电容２５μＦ，电场强度１０．０ＫＶ／ｃｍ，脉冲控制器设定２００。 Ｋ１０４１和 Ｔ５２１最高转化效率分别达２．０１×１０和１．１９ ×１０转化子／μｇ ＤＮＡ。此外，研究发现Ｔ５２１和Ｋ１０４１中存在着ＤＮＡ的限制／修饰系统。
Transformation of shuttle plasmids into Bacillus stearothermophilus K1041 and T521 by electroporation was carried out. K1041 cells for transformation were prepared from late-logarithmic growth phage, T521 cells prepared after growth on LB plate at 50℃ overnight. The cell concentration was about 5-7 × 10cells/mL. The electroporation conditions were to set capacitor at 25μF, electrical field strength at 10.0KV / cm and the pulse controller at 200 ohms. The maximum transformation efficiencies of K1041 and T521 were up to 2.01 × 104 and 1.19 × 102 transformants/μg DNA, respechvely. In addition,evidence indicated that T521 and K1041 possessed DNA restriction and modification systems.
出处
《微生物学通报》
CAS
CSCD
北大核心
2000年第2期85-89,共5页
Microbiology China
基金
国家自然科学基金!No.39230020
关键词
嗜热脂肪芽胞杆菌
质粒
电击转化
B. stearothermophilus, Plasmid, Electrotransformation
