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发明公开
CN105126092A 一种以壳寡糖为佐剂负载大肠杆菌菌毛黏附素的鸭疫里默氏菌菌影疫苗的制备方法及其应用 无效 - 驳回

一种以壳寡糖为佐剂负载大肠杆菌菌毛黏附素的鸭疫里默氏菌菌影疫苗的制备方法及其应用
摘要：
本发明公开了一种以壳寡糖为佐剂负载大肠杆菌菌毛黏附素的鸭疫里默氏菌菌影疫苗的制备方法及应用。通过将从鸭源大肠杆菌克隆得到的编码fimA黏附素亚单位的pilA基因与噬菌体PhiX174的E基因，通过编码柔性连接臂(GLy4Ser)3的Linker连接，获得pilA、E的串联基因pilA-E，将该串联基因pilA-E与温控表达载体pBV220连接，构建温控表达载体pBV-pilA-E，将所述pBV-pilA-E电转化为鸭疫里默氏菌感受态细胞，并筛选阳性克隆体从而得到鸭疫里默氏菌工程菌，离心分离菌体，得到鸭疫里默氏菌菌影，将壳寡糖配的水溶液加入到菌检无活菌的鸭疫里默氏菌菌泥中，得到鸭疫里默氏菌菌影壳寡糖佐剂疫苗。本发明所提供的鸭疫里默氏菌菌影疫苗，可以通过饮水免疫，应用方便，适应规模化养殖。
摘要（英）：

The invention discloses a preparation method for a riemerella anatipestifer ghost vaccine with chitosan oligosaccharide as adjuvant load escherichia coli pili adhesion and application thereof. The method comprises the steps that a gene pilA of a code fimA adhesion subunit and a gene E of a phage PhiX174 obtained through duck-derived escherichia coli cloning are connected with a Linker of a code flexible connecting arm (GLy4Ser)3 to obtain a serial gene pilA-E of pilA,E, the serial gene pilA-E is connected with a temperature control expression vector pBV220 to construct a temperature control expression vector pBV-pilA-E, the pBV-pilA-E is electrically converted into a riemerella anatipestifer competent cell, and a positive clone body is screened out, so that riemerella anatipestifer enginnering bacteria are obtained, centrifugal separation is performed on the bacteria to obtain a riemerella anatipestifer ghost, and a water colution prepared from chitosan oligosaccharide is added into riemerella anatipestifer sludge with no living bacteria through bacterium testing to obtain the riemerella anatipestifer ghost chitosan oligosaccharide adjuvant vaccine. The riemerella anatipestifer ghost vaccine can achieve immunity through drinking water, and is convenient to apply and suitable for large scale breeding.

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