Optimum Conditions for Electric Pulse-mediated Gene Transfer to Bacillus subtilis Cells
Optimum Conditions for Electric Pulse-mediated Gene Transfer to Bacillus subtilis Cells
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电脉冲介导的基因转移至枯草芽孢杆菌细胞的最佳条件
DOI：
10.1271/bbb1961.53.2441
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
H. Kimoto
中科院分区：
文献类型：
--
作者：
H. Kusaoke;Y. Hayashi;Yasuhiro Kadowaki;H. Kimoto
结果表明，脉冲电场不仅能将质粒DNA（pUB 110）导入枯草芽孢杆菌的原生质体，而且能将质粒DNA导入完整的细胞。的变换，B。用原生质体转化枯草芽孢杆菌，在初始电场强度为7 kV/cm，脉冲时间为50 μ sec时，转化效率为2.5 × 104/μg DNA。完整B的均匀转化。在初始电场强度为16 kV/cm、脉冲时间为400 μsec的条件下，转化效率最高可达1.5 × 103个转化子/μ gDNA。原生质体和完整细胞的细胞存活率分别约为100%和30%，在转化过程的最佳条件下。在凝胶电泳分析中，从含有pUB 110的转化体中分离的质粒DNA与pBU 110的真实制备物无法区分。
It was found that plasmid DNA (pUB 110) can be introduced into not only protoplasts but also intact cells of Bacillus subtilis by electric field pulses. The transformation of, B. subtilis using protoplasts results in an efficiency of 2.5 × 104 transformants per μg of DNA, with a single pulse of 50 jisec with an initial electric field strength of 7kV/cm. Even transformation of intact B. subtilis cells results in a maximum efficiency of 1.5 × 103 transformants per μg DNA, with a single pulse of 400 μsec with an initial electric field strength of 16kV/cm. The cell survival of protoplasts and intact cells was approximately 100% and 30%, respectively, under the conditions found to be optimal for the transformation process. Plasmid DNA isolated from pUB 110 containing transformants was indistinguishable from authentic preparations of pBU 110 on gel electrophoretic analysis.