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目的探讨制备脂质体包裹重组SEF21疫苗,并评价其在预防肠炎沙门菌(S.enteritidis)感染中的作用。方法利用PCR获得SEF21基因,并连接至pET-28a(+)载体。将pET-28a(+)-SEF21在BL21(DE3)大肠埃希菌中表达,通过镍层析柱纯化高表达的rSEF21蛋白。制备脂质体包裹rSEF21疫苗,并对鸡进行2次免疫,然后利用S.enteritidis进行攻毒实验。ELISA检测血清以及肠内容物中的抗体效价。结果所有被免疫鸡的血清及肠黏液中产生了高效价的IgG和IgA抗体。脂质体包裹rSEF21所免疫的鸡的粪便样本中S.enteritidis数量明显下降。结论口服脂质体包裹的重组SEF21蛋白疫苗能有效保护鸡对抗S.enteritidis感染。
Objective To investigate the preparation of liposome-encapsulated recombinant SEF21 vaccine and evaluate its role in the prevention of S. enteritidis infection. Methods The SEF21 gene was obtained by PCR and ligated into pET-28a (+) vector. PET-28a (+) - SEF21 was expressed in BL21 (DE3) Escherichia coli and the highly expressed rSEF21 protein was purified by nickel chromatography. The liposome-encapsulated rSEF21 vaccine was prepared and the chickens were immunized twice, then challenged with S.enteritidis. ELISA to detect antibody titer in serum and intestinal contents. Results High titer IgG and IgA antibodies were produced in serum and intestinal mucus of all immunized chickens. The number of S. enteritidis in stool samples of chickens immunized with liposome-encapsulated rSEF21 was significantly decreased. Conclusions Oral liposome-encapsulated recombinant SEF21 protein vaccine can effectively protect chickens against S.enteritidis infection.