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《Chinese Journal of Zoonoses》 2010-07
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Generation and characterization of enterohemorrhagic Escherichia coli O157 ghosts

LIU Shuang1,2,LIU Jun2,3,ZHOU Bo2,3,WANG Wen-dong1,3,LI Peng2,3,ZHU Ling-wei2,3,JI Xue2,3,FENG Shu-zhang1,2,3(1.College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China;2.Institute of Veterinary Science,Academy of Military Medical Sciences,Changchun 130062,China 3.Key Laboratory of Jilin province for Zoonosis Prerention and Contral,Changchun 130062,China)  
Enterohemorrhagic Escherichia coli(EHEC)O157∶H7 is an important food borne pathogen capable of causing non-bloody or bloody diarrhea,hemorrhagic colitis(HC)and potentially fatal hemolytic uremic syndrome(HUS).Many efforts have been made by several research groups to develop vaccine.However there is not any vaccine could be used in clinic.Bacterial ghosts are nonliving bacterial cell envelopes devoid of cytoplasmic contents while maintaining their cellular morphology and native surface antigenic structures.Therefore,bacterial ghosts could be the excellent vaccine candidates.In the present study,lysis gene E was amplified by PCR from phage PhiX174 and inserted into pBV220 vector downstream of the λPL/PR-cI857 regulatory system to form the temperature-sensitive bacteriolysis plasmid(pBV220::E).Plasmid pBV220::E was transformed into O157∶H7 and grown at 28℃ until mid log-phase,followed by incubation at 42℃ to induce the expression of gene E and generate O157∶H7 bacterial ghosts.The OD600 value of culture began to decline in 40 min after the temperature shift from 28℃ to 42℃.The lysis rate of culture was 98.4%.The O157∶H7 ghosts were shown to be intact cells under electron microscope,with contents released to extracellular region.In conclusion,the O157∶H7 ghosts were successfully generated with lysis gene E,which laid foundation for future development of bacterial ghost vaccine against EHEC O157∶H7 infection.
【Fund】: 国家支撑计划(2007BAB55B05);; 国家自然科学基金(30270985)联合资助
【CateGory Index】: R378
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