Method for preparing recombinant spore for surface display of prawn white spot syndrome virus Vp28
A technology of white spot syndrome and surface display, applied in the direction of botanical equipment and methods, microorganism-based methods, biochemical equipment and methods, etc., to achieve good stability
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Embodiment 1
[0041] Preparation of recombinant spores of Bacillus subtilis displaying Vp28 on the surface using CotC as carrier protein
[0042] 1. Molecular Biology Operations
[0043] 1.1 Extraction of Bacillus subtilis chromosome
[0044] Collect 10mL Bacillus subtilis168 (trp - ) (Bacillus Genetic Stock Center, Department of Biochemistry The Ohio State University 484 West Twelfth Avenue Columbus, Ohio 43210, USA) culture, add 0.5ml TE to suspend the precipitate. Add 30μl lysozyme (100mg / ml) to each microcentrifuge tube, react at 37°C for 1 hour; add 50μl SDS (10%) and 20μl proteinase K (20mg / ml), shake evenly, react at 37°C for 2 hours, add etc. Take the supernatant after extraction with phenol / chloroform, add 2 times the volume of ethanol, precipitate the DNA for more than 2 hours at room temperature, centrifuge at 12,000g for 10 minutes, discard the supernatant and wash the DNA precipitate with 500μl of 75% ethanol to Remove inorganic salt ions. After the DNA precipitate is dry, ...
Embodiment 2
[0074] Preparation of recombinant spores of Bacillus subtilis displaying Vp28 on the surface using CotG as carrier protein
[0075] 1. Molecular Biology Operations
[0076] 1.1 Extraction of virus chromosome
[0077] Concrete operation method is with the extraction of embodiment 1 of the present invention, 1.1 virus chromosome
[0078] 1.2 Extraction of Bacillus subtilis chromosome
[0079] Concrete operation method is with the extraction of the embodiment of the present invention 1,1.2 Bacillus subtilis chromosome
[0080] 1.3 Molecular biology techniques
[0081] Concrete operation method is with the embodiment of the present invention 1,1.3 molecular biology technology
[0082] 1.4PCR amplification
[0083] Concrete operation method is with the embodiment of the present invention 1,1.4 molecular biology technology
[0084] 2. Plasmid construction
[0085] 2.1 Basic plasmid construction
[0086] Concrete operation method is the same as embodiment 1, the construction ...
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