Tagged swine fever virus e2 protein recombinant baculovirus inactivated vaccine
A technology of recombinant baculovirus and swine fever virus, applied in the field of vaccines, can solve problems affecting vaccine safety, contaminated cattle viral diarrhea, and uneven quality of antigens
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0071] Embodiment 1 Swine fever virus E2 gene optimization, mutation and synthesis
[0072]The published E2 sequences of swine fever virus and the E2 sequences of new strains popular in recent years were downloaded from Genebank for reference, and the E2 gene of strain C was optimized based on the optimized E2 gene and the reference strains of each subgroup. The alignment was performed, and the results showed that the optimized E2 gene was identified as group 1.1. At the same time, the optimized E2 gene sequence epitope "TAVSPTTLR" was mutated to "ATVSPTTRL, TASVPTTLR, TAVSTPTLR, TAVSPTTRL, TAPVSTTRL or TASPVTTRL", and the obtained 6 optimized mutated sequences were handed over to Gene Company for synthesis.
Embodiment 2
[0073] The construction of embodiment 2 transfer vector
[0074] The test method is referred to the Invitrogen operation manual, and the specific operation is described as follows.
[0075] formulate The clone reaction mixture has the following components: 0.5-4µl of synthesized E2 nucleotides, 1µl of salt solution (1.2mol / LNaCl, 0.06mol / LMgCl2)), 1µl carrier, and double-distilled water was added to bring the total volume to 6 μl. The steps of the cloning reaction are as follows: tap the wall of the centrifuge tube to make the reaction mixture evenly mixed, leave it at room temperature (22-23°C) for 15 minutes, and place the reaction tube on ice. Take 2μl Add the clone reaction solution to the competent cells, tap the tube wall to mix evenly, put it on ice for 30 minutes, then place it in a 42°C water bath for 30 seconds for heat shock, and immediately put the reaction tube back on ice. Then, 250 μl of the warmed SOC culture medium was added to the reaction tube, and i...
Embodiment 3
[0077] Example 3 Recombination of transfer vector and linear baculovirus
[0078] The extracted recombinant vector was used to measure the nucleic acid concentration of the recombinant vector with a spectrophotometer at a wavelength of 260 nm. The LR recombination reaction was carried out according to the operation manual of "BaculoDirectTM Baculovirμs Expression System" provided by Invitrogen. The specific operations are as follows: aseptically transfer 1 μl of recombinant transfer vector (100-300 ng), 4 μl of 5×LR The reaction buffer and 1 μl of double-distilled water were added to 10 μl of BaculoDirectTM linear DNA (300 ng), respectively. Remove LR from -70°C freezer The enzyme mix was placed on ice and warmed for 2 minutes, shaken twice, 2 seconds each time, and 4 μl LR was taken. Add the enzyme mix to the above mixture, tap the tube wall several times to mix evenly, do not shake vigorously to avoid baculovirus DNA breakage. After 18 hours at 25°C, 2 µl of proteinas...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More