Construction of an alpha toxin gene knockout mutant of Clostridium perfringens type A by use of a mobile group II intron
Construction of an alpha toxin gene knockout mutant of Clostridium perfringens type A by use of a mobile group II intron
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DOI：
10.1128/aem.71.11.7542-7547.2005
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发表时间：
2005-11-01
影响因子：
4.4
通讯作者：
Gupta, P
中科院分区：
文献类型：
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作者：
Chen, Y;McClane, BA;Gupta, P
在开发作为安全疫苗载体的产气荚膜梭菌时，细菌染色体中的α毒素基因（plc）必须永久失活。通过传统的突变方法破坏产气荚膜梭菌中的基因是非常困难的。因此，我们开发了一种新的策略，使用基于第11组内含子的Target-Tron技术在C.产气荚膜杆菌ATCC 3624。蛋白质印迹分析显示在plc突变体的培养上清液中没有产生α毒素蛋白。该技术具有位点特异性、插入频率高、不引入抗生素抗性基因等优点，可为其它C.产气荚膜杆菌突变体。
In developing Clostridium perfringens as a safe vaccine vector, the alpha toxin gene (plc) in the bacterial chromosome must be permanently inactivated. Disrupting genes in C perfringens by traditional mutagenesis methods is very difficult. Therefore, we developed a new strategy using group 11 intron-based Target-Tron technology to inactivate the plc gene in C. perfringens ATCC 3624. Western blot analysis showed no production of alpha toxin protein in the culture supernatant of the plc mutant. Advantages of this technology, such as site specificity, relatively high frequency of insertion, and introduction of no antibiotic resistance genes into the chromosome, could facilitate construction of other C. perfringens mutants.