Prepn and usage of extracellular product subunit vaccine of seawater fish morbid vibrio
A subunit vaccine and extracellular product technology is applied in the field of preparation of marine fish pathogenic Vibrio extracellular product subunit vaccine, which can solve the problems of large dosage, easy rebound, poor immune effect, etc., and achieve immune specificity. Stable and easy-to-use effects
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Embodiment 1
[0035] Vibrio alginolyticus and Vibrio harveyi were first activated with trypticase soytone liquid medium (TSB), cultured at 28°C with shaking at 150r / min for 18h, and then 1mL of the bacterial solution was spread on sterile cellophane The tryptone soy agar plate medium (TSA) containing 2% NaCl and the initial pH of 7.2 was cultured statically at 28° C. for 18 h. Then, extracellular products of Vibrio alginolyticus and Vibrio harveyi were extracted respectively. The method is as follows: wash the bacterial liquid with PBS of pH 7.2, and centrifuge at 10,000 r / min for 30 min at 4°C. The supernatant was filtered through a microporous membrane with a pore size of 0.22 μm to obtain Vibrio extracellular products (ECP). Determine the protein concentration in the samples with a UV spectrophotometer. Next, the extracellular product subunit vaccines of Vibrio alginolyticus and Vibrio harveyi were prepared respectively. The method is as follows: add 0.1% formalin to the extracellular...
Embodiment 2
[0037] Vibrio alginolyticus, Vibrio harveyi and Vibrio vulnificus were first activated with trypticase soytone liquid medium (TSB), cultured at 28°C with shaking at 150r / min for 18h, and then 1mL of the bacterial solution was spread on the shop floor respectively. Trypticase soy agar plate medium (TSA) containing 2% NaCl and an initial pH of 7.2 with sterile cellophane was cultured statically at 28°C for 18 hours. Then, extracellular products of Vibrio alginolyticus, Vibrio harveyi and Vibrio vulnificus were extracted respectively. The method is as follows: wash the bacterial liquid with PBS of pH 7.2, and centrifuge at 10,000 r / min for 30 min at 4°C. The supernatant was filtered through a microporous membrane with a pore size of 0.22 μm to obtain Vibrio extracellular products (ECP). Determine the protein concentration in the samples with a UV spectrophotometer.
[0038] Next, the extracellular product subunit vaccines of Vibrio alginolyticus, Vibrio harveyi and Vibrio vulni...
Embodiment 3
[0040] Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus and Vibrio anguillarum were first activated with trypticase soytone liquid medium (TSB), cultured at 28°C with shaking at 150r / min for 18h, and then 1 mL of the bacterial liquid was taken separately Spread on tryptone soy agar plate medium (TSA) containing 2% NaCl and initial pH 7.2 covered with sterile cellophane, and cultured statically at 28°C for 18h. Then, the extracellular products of Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus and Vibrio anguillarum were extracted respectively. The method is as follows: wash the bacterial liquid with PBS of pH 7.2, and centrifuge at 10,000 r / min for 30 min at 4°C. The supernatant was filtered through a microporous membrane with a pore size of 0.22 μm to obtain Vibrio extracellular products (ECP). Determine the protein concentration in the samples with a UV spectrophotometer. Next, the extracellular product subunit vaccines of Vibrio alginolyticus, Vibrio harvey...
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