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Outline
Infectious bursal disease （IBD） severely restricts the economic development of the poultry industry. Therefore， the development of safe and efficient IBD vaccines has become a key topic of considerable interest. To explore the feasibility of using Eimeria tenella as a live vector for the expression and delivery of infectious bursal disease virus （IBDV） antigens， this study constructed a genetically modified strain of E. tenella （Et-VP2） expressing the VP2 protein of the IBDV. An expression plasmid was constructed， digested with enzymes， and then transfected into E. tenella using nuclear transfection. Flow cytometry was utilized to sort and select positive strains expressing the red fluorescent reporter gene （mCherry）， and the expression of the VP2 gene was identified at both the genetic and protein levels. Results showed that after three passages in vivo in chicks， the transgenic E. tenella expressing VP2 protein had a positive rate of fluorescence exceeding 95%， confirming successful integration and stable expression of the foreign gene at both the genetic and protein levels （with fluorescence rates reaching 95% in the offspring population）. Indirect immunofluorescence results indicated that the fusion protein （Mic2-linker-VP2） was localized at the apical end of the sporozoites of E. tenella. This study successfully established a stable transfection system for E. tenella， laying the foundation for subsequent research on the immune protection of genetically modified strains against infectious bursal disease.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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