Preparation of immuno-stimulation composition for hemolycin in monad
A technology for Aeromonas hydrophila and immunostimulation, applied in the fields of molecular biology and immunology, can solve the problems of Aeromonas hydrophila complex antigen structure, weak immunogenicity, inconvenient preparation of combined vaccines, etc., to achieve Strong immune protection, stable immune effect, and remarkable synergistic effect
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Embodiment 1
[0063] 1. Required materials
[0064] (1) Main reagents
[0065] Quil A was purchased from Beijing Zhongjian Weikang Technology Co., Ltd.; Mega-10 was purchased from Huamei Bioengineering Company.
[0066] (2) Strains
[0067] Aeromonas hydrophila L-316 was isolated and identified by the Fish Disease Center of the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, and its β-hemA recombinant bacteria were constructed.
[0068] (3) Experimental animals
[0069] Eel, purchased from Changle Eel Farm in Fujian Province, size: about 200g.
[0070] Two, the preparation technology of the immunostimulatory complex of hemolysin in Aeromonas hydrophila, has the following steps:
[0071] 1. Extract the recombinant bacteria of Aeromonas hydrophila L316,
[0072] 2. In the culture medium, the bacterial strain is expanded and cultivated to obtain a culture solution,
[0073] (1) inoculate the bacterial strain on common agar medium,
[0074]...
Embodiment 2
[0125] The citric acid in the step of the preparation process of the immunostimulatory complex of hemolysin in Aeromonas hydrophila of Example 1 is replaced with the glycine buffer solution of pH2.5 for preparing the immunostimulatory complex of Aeromonas hydrophila thing;
[0126] Concentrate the toxin to 1 mg / ml and add glycine buffer solution with pH 2.5 for 1 hour;
[0127] Add Quil A adjuvant to a final concentration of 0.05%;
[0128] The remaining experimental materials and experimental procedures are the same as in Example 1. Experimental results: the 40ug dose group has the best effect, and the protection rate reaches 100%, followed by the 80ug dose group. It can be seen that the toxin protein is treated with glycine and citric acid. The effect of the immunostimulatory complex for humoral immunity to immune receptors is basically the same.
[0129] Best practice:
[0130] The steps of the preparation process of the immunostimulatory complex of hemolysin in Aeromona...
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