Construction of eivpP in-frame deletion of pathogenic bacterium Edwardsiella ictaluri
GUO Yi, YANG Xiao-yu, NI Ping, WANG Yi-ting, GUO Si-cong, MA Hong-li, ZHAO Xiao-ran, YE Shi-gen
Key Laboratory of Mariculture & Stock Enhancement in North China’s Sea, Ministry of Agriculture and Rural Affairs, Dalian Key Laboratory of Marine Animal Disease Prevention and Control, Dalian Ocean University, Dalian 116023, China
Abstract: In order to study the interaction between EivpP and other secreted proteins of Edwardsiellaictaluri type Ⅵ secretion system (T6SS) and its mechanism in the pathogenesis of bacteria, the fusion fragment containing the upstream and downstream homologous arms for the eivpP gene deletion was prepared by fusion PCR and successfully cloned into the suicide plasmid pDM4.The recombinant suicide plasmid was transformed into Escherichacoli S17-1(pir), co-cultured with E.ictaluri wild strain ATCC33202, and E.ictalurieivpP gene deletion mutation positive clone was obtained by homologous recombination and sucrose reverse screening.The results of PCR identification showed that the eivpP gene was not detected in the mutant strain, that is, the eivpP gene was successfully knocked out.The findings provide data with research of the role of eivpP gene in the pathogenesis of E.ictaluri, and provide a theoretical basis for the development of E.ictaluri vaccine as well.
2019, Vol. 34 
