摘要
旨在制备禽致病性大肠杆菌菌蜕,并优化制备条件。将温控型表达质粒pBV221-E通过电击的方式,电转化入12株禽致病性大肠杆菌分离株中,通过改变温度诱导E基因表达来裂解细菌制备菌蜕;同时探究了不同起始诱导A_(600)值(0.4、0.5、0.6)对裂解效率的影响,优化制备条件提高裂解效率,并评估禽致病性大肠杆菌菌蜕的免疫保护力。结果:不同菌株的裂解效率有较大差异,在起始诱导A_(600)值为0.5时,禽致病性大肠杆菌D78菌蜕的裂解效率最高,可达约98.00%;经透射电镜观察发现,制备获得的菌蜕表面有明显孔道,细胞质从孔道溢出,菌体电子密度下降;免疫效力评估试验显示,SPF鸡经肌肉注射D78菌蜕疫苗,二免14 d后,通过气囊注射攻毒,D78菌蜕疫苗的免疫保护率为66.67%,优于D78甲醛灭活疫苗。研究结果为新型禽致病性大肠杆菌疫苗的研发奠定了基础。
In order to prepare a bacterial ghost of avian pathogenic Escherichia coli and to optimize the preparation conditions,the temperature-controlled expression plasmid pBV221-E was electro-transformed into 12 avian pathogenic Escherichia coli strains by electric shock.The E gene expression was induced by changing the temperature to lyse the bacteria to prepare the bacterial ghost.At the same time,the effects of different initial induction A_(600) values(0.4,0.5,0.6)on the lysis efficiency were explored,the preparation conditions were optimized to improve the lysis efficiency,and the immune protection of avian pathogenic Escherichia coli ghosts was evaluated.The results showed that the lysis efficiency of different strains evidently varied.When the initial induction A_(600) value was 0.5,the lysis efficiency of avian pathogenic Escherichia coli D78 ghosts was the highest,reaching about 98.00%.The transmission electron microscopy showed that there were obvious pores in the surface of the prepared bacterial ghost,and the cytoplasm overflowed from the pores,and the electron density of the bacteria decreased.The immune efficacy evaluation test showed that 14 days after the SPF chickens were intramuscularly injected with D78 bacterial ghost vaccine,the immune protection rate of the D78 bacterial ghost vaccine was 66.67%,which was better than that of the D78 formaldehyde inactivated vaccine.The present study laid a foundation for development of new avian pathogenic Escherichia coli vaccines.
作者
刘思琪
郭梦娇
吴艳涛
袁橙
LIU Siqi;GUO Mengjiao;WU Yantao;YUAN Cheng(College of Veterinary Medicine of Jiangsu Agricultural and Animal Husbandry Vocational College,Taizhou 225300,China;School of Veterinary Medicine,Yangzhou University,Yangzhou 225000,China)
出处
《畜牧与兽医》
北大核心
2025年第7期97-102,共6页
Animal Husbandry & Veterinary Medicine
基金
江苏省高等学校自然科学研究面上项目(21KJB230007)
泰州市科技支撑计划(农业)项目(TN202118)
江苏高校“青蓝工程”项目(苏教师函[2024]14号)。
关键词
菌蜕
禽致病性大肠杆菌
E基因
裂解效率
bacterial ghost
avian pathogenic Escherichia coli
gene E
lysis efficiency
