| 用十二烷基硫酸钠消除枯草芽孢杆菌中的质粒 |
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| 引用本文： | 娄恺,班睿,赵学明.用十二烷基硫酸钠消除枯草芽孢杆菌中的质粒[J].天津大学学报(英文版),2002,8(3):148-151. |
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| 作者姓名： | 娄恺 班睿 赵学明 |
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| 作者单位： | 1. 天津大学化工学院,天津300072;新疆农业科学院微生物研究所,乌鲁木齐830000
2. 天津大学化工学院,天津300072 |
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| 基金项目： | SupportedbyKeyProjectofNaturalScienceFoundationofChina(No .2 0 0 360 1 0 ) |
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| 摘 要： | 为获得宿主菌 ,研究了十二烷基硫酸钠 (SDS)对枯草芽孢杆菌中质粒的消除 .将过夜培养的枯草芽孢杆菌2 4/pMX45接种于含SDS( 0— 0 .0 0 8% )的LB培养基中 ,当SDS浓度 (w /v)大于或等于 0 .0 0 6 %时 ,菌体不能生长 .SDS的亚致死浓度为 0 .0 0 5 % ,其致死率达 99% .菌液稀释后涂LB平板 ,再随机挑选单菌落至抗性平板 ,检测由质粒编码的红霉素抗性是否丢失 .氯化铯 溴化乙锭梯度离心及质粒DNA的电泳图像证实了 2 4/pMX45的衍生菌株A7中的质粒已完全被消除 .A7延迟期较短 ,并且细胞浓度高于 2 4/pMX45 .用SDS处理 8h后 ,2 4/pMX45的遗传标记开始丢失 ,消除率持续增高至 2 2h ,随之消除质粒的菌体量保持恒定 .在未经SDS处理的对照实验中 ,相同条件下培养 2 4h及 48h后 ,没有发现质粒自然丢失的现象 ,因此SDS能消除枯草芽孢杆菌中的质粒
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| 关 键 词： | 枯草芽孢杆菌 质粒 消除 十二烷基硫酸钠 |
Curing of the Bacillus subtilis Plasmid Using Sodium Dodecyl Sulfate |
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| LOU Kai,BAN Rui,ZHAO Xue-ming.Curing of the Bacillus subtilis Plasmid Using Sodium Dodecyl Sulfate[J].Transactions of Tianjin University,2002,8(3):148-151. |
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| Authors: | LOU Kai BAN Rui ZHAO Xue-ming |
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| Abstract: | Curing of Bacillus subtilis plasmid using sodium dodecyl sulfate (SDS)was studied in order to obtain a host strain. An overnight culture of Bacillus subtilis 24/pMX45 was used to inoculate were randomly picked to a selective agar medium by tooth to screen for loss of plasmid-encoded erythomycin resistance. CsCl-EtBr gradient centrifugation and plasmid DNA profile demonstrated that plasmid-cured derivative A7 has completely lost its plasmid. A7 had a shorter lag, and its cell concentration was consistently higher than that of the 24/pMX45. Elimination of the plasmid was first observed after 24/pMX45 had been treated with SDS for 8 h. The percent elimination then continued to increase until about 22 h, after which the fraction of cured cell in the population remained constant. Plasmid cured cell numbers were measured in a separate control culture of 24/pMX45 untreated by SDS. No spontaneous loss of pMX45 was observed after 24/pMX45 were incubated for 24 h and 48 h with shaking at 37 ℃.These results suggested that SDS can be used as curing agent to eliminate the plasmid of Bacillus subtilis. |
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| Keywords: | Bacillus subtilis plasmid curing sodium dodecyl sulfate |
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