Electrotransformation of Foodborne Pathogen Cronobacter sakazakii by a Simple Method

Wei Chen,Zhimeng Liu, Wenjie Sun,Siqi Li, Jiajia Liu,Weiping Huo, Jia Jia, Wenyan Shen,Yuanyuan Wang,Gukui Chen

FOODBORNE PATHOGENS AND DISEASE(2024)

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摘要
Cronobacter sakazakii is an opportunistic foodborne pathogen that mainly infects infants and immunocompromised people, with a high mortality rate. However, the efficient transformation method of this bacterium has not been systematically reported. In this study, we developed a fast and efficient transformation method for C. sakazakii by cold sucrose treatment. Compared with CaCl2 or glycerol treatment, the transformation efficiency of this method is significantly high when bacteria were cultured overnight at 42(degrees)C before cold sucrose treatment. Furthermore, applying this method, we successfully knocked out the pppA gene by direct electroporation. Collectively, our study provides a simple, time-saving, and efficient method for competent cell preparation of C. sakazakii, which is conducive to the further research of C. sakazakii.
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关键词
Cronobacter sakazakii,electrotransformation,sucrose,temperature
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