制备禽致病性大肠杆菌菌蜕的三种方法比较

发布时间：2018-11-27 08:29

【摘要】：菌蜕(Bacterial ghosts)是一种只含有细菌内、外膜结构的细菌空壳,可作为新型疫苗和传递载体。本研究通过3种方式制备禽致病性大肠杆菌(Avian pathogenicity Escherichia coli,APEC)分离株DE17的菌蜕,评价不同的菌蜕制备方法。结果表明,利用噬菌体Phi X174的裂解基因E构建的溶菌质粒pBV220-E制备DE17菌蜕,对DE17菌株裂解率可达99.9%,扫描电镜观测结果表明,在DE17两端或中部形成可见的跨膜孔道,呈现典型的菌蜕结构。利用合成的细胞穿透肽MAP(KLALKLALKALKAALKLA)作用于DE17制备菌蜕,结果表明,10μmol/L的MAP可实现对OD_(600)=0.1的DE17完全灭活,扫描电镜虽未看到明显的跨膜孔道,但细菌的膜结构呈现沟壑状,而构建的可表达MAP的溶菌质粒pBV220-MAP并不能实现对DE17的裂解作用。本研究通过比较分析不同APEC菌蜕的制备方式,为进一步研究菌蜕疫苗和提高菌蜕疫苗的生物安全性提供参考。
[Abstract]:Bacterial slough (Bacterial ghosts) is a kind of bacterial shell containing only internal and outer membrane structure, which can be used as a new vaccine and delivery carrier. The purpose of this study was to prepare the DE17 slough of avian pathogenic Escherichia coli (Avian pathogenicity Escherichia coli,APEC) strain by three methods and evaluate the different preparation methods. The results showed that the lysogenic plasmid pBV220-E constructed from the lytic gene E of phage Phi X174 was used to prepare DE17 slough. The cleavage rate of DE17 strain could reach 99.9%. The results of scanning electron microscope showed that transmembrane channels could be formed at the two ends or the middle of DE17. A typical mycelial slough structure is presented. The cell penetrating peptide (MAP (KLALKLALKALKAALKLA) was used in the preparation of DE17. The results showed that 10 渭 mol/L MAP could completely inactivate the DE17 of OD_ (600) = 0.1, and no obvious transmembrane pore was observed under SEM. However, the membrane structure of bacteria was gully, and the lysogenic plasmid pBV220-MAP, which could express MAP, could not realize the cleavage of DE17. In this study, the preparation methods of different APEC slough were compared and analyzed in order to provide a reference for further research on the microbial slough vaccine and the improvement of the biological safety of the mycelium slough vaccine.
【作者单位】：西南大学动物科学学院;中国农业科学院上海兽医研究所;扬州大学动物科学与技术学院江苏省人畜共患病学重点实验室;
【基金】：上海市科技兴农重点攻关项目(No.2015HNG1-9) 国家自然科学基金(No.31370045) 江苏省人兽共患病学重点实验室项目(No.R1510)资助~~
【分类号】：S859.797

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