Chicken subunit four-combination vaccine and preparation and application thereof
A vaccine and chicken embryo technology, applied in the field of avian vaccine manufacturing, can solve the problems of short immunization period, easy to cause respiratory side effects, and interference with laboratory diagnosis of virus isolation, etc.
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[0014] The main structural protein VP2 protein of IBDV is the most important host protective antigen. The present invention amplifies the super-virulent VP2 gene of IBD by PCR method, clones it on the pET-28a vector, transforms BL21(DE3), and induces it with IPTG After the VP2 protein is expressed, the expressing cells are homogenized under high pressure and then centrifuged to obtain the supernatant, which is the VP2 protein solution.
[0015] Newcastle disease virus Clone30 strain was inoculated into 10-day-old SPF embryos, and the chicken embryo liquid cultured for 48-96 hours was collected. The virus liquid was concentrated by ultrafiltration and concentration, and 0.1% formaldehyde was added to the final concentration.
[0016] Chicken infectious bronchitis virus M42 strain was used to inoculate 10-11-day-old SPF embryos, and the chicken embryo fluid cultured for 24-48 hours was collected. The virus was concentrated by ultrafiltration and concentration, and 0.1% formaldehy...
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