Eimeria acervulina nano-subunit vaccine, preparation method and application thereof
A technology of Eimeria and heap-type, which is applied in the field of Eimeria heap-type nano-subunit vaccine and its preparation, can solve the problems such as the application research report of PLGA nano-subunit vaccine, and achieve the effect of immune protection The effect of enhancement and embedding rate improvement
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Embodiment 1
[0029] Example 1. Expression and purification of Eimeria acervulina recombinant protein EaMIC3
[0030] 1. Preparation of genetically engineered bacteria
[0031] The E. acervulina pET-32a-EaMIC3 ORF (Zhang Zhenchao, the role of Eimeria acerciformis microwire protein in the specificity of invasion site. Nanjing Agricultural University PhD dissertation, 2016) was transformed into E.coli BL21 receptor. Cells, and get bacteria containing pET-32a-EaMIC3 ORF recombinant expression plasmid.
[0032] 2. Expression and purification of Eimeria acervulina recombinant protein EmMIC3
[0033] Bacteria containing pET-32a-EaMIC3 ORF recombinant expression plasmid were inoculated into LB liquid medium at a volume ratio of 1:100, and cultivated to OD at 37℃, 200r / min 600 When it is 0.4-0.6, add IPTG with a final concentration of 1mmol / L to induce expression. Purify the EMIC3 recombinant protein in accordance with the GE protein purification kit instructions, and subject the collected protein samples...
Embodiment 2
[0036] Example 2. Immune protection test of Eimeria acervulina recombinant protein subunit vaccine and nano subunit vaccine 1. Experiment design
[0037] 0-day-old Hailan white chicks are raised in a strictly disinfected, coccidia-free environment, and they are free to drink and eat. The 14-day-old chickens were weighed one by one, and overweight and underweight individuals were eliminated. Randomly grouped and adjusted the average weight between the test groups so that the average weight was close to the same, 15 birds in each group; recombinant proteins EaMIC3 and pET- were used respectively. The PLGA-EaMIC3 and PLGA-pET-32a-tagged proteins are nano subunit vaccines prepared by encapsulating recombinant proteins with 32a-tagged protein, PLGA, and PLGA nanomaterials to immunize chickens by intramuscular injection into the leg. The immunization dose is 100μg, and it is administered at 21 days of age. Booster immunity, oral infection of fresh E.acervulina sporulated oocysts 10×10 ...
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