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In this study， we aimed to develop recombinant PRV rPRV-AH-gI-/gE-/gM-/gD+ with double gD and evaluate its immune efficacy. The recombinant virus rPRV-AH-gI-/gE-/gM-/gD+ was constructed by deleting gM gene and inserting gD expression cassette at the deficient region. Four-week-old Kunming mice were immunized twice with 107.0 TCID50 inactivated rPRV-AH-gI-/gE-， rPRV-AH-gI-/gE-/gM-， rPRV-AH-gI-/gE-/gM-/gD+ at an interval of 3 weeks. Commercial vaccine and DMEM were as the control group. After immunization， regular blood was collected， neutralizing antibody levels in sera were detected by neutralization test， and total antibody levels were detected by ELISA. At 6 weeks of first immunization， mice were challenged with 105.5TCID50 PRV-AH. The inserted gD gene was expressed with the molecular weight of 53 ku in the rPRV-AH-gI-/gE-/gM-/gD+. The neutralizing and total IgG antibody levels of rPRV-AH-gI-/gE-/gM-/gD+ group were significantly higher than those of rPRV-AH-gI-/gE and rPRV-AH-gI-/gE-/gM- at 4 and 6 weeks after first immunization （P<0.01）. However， compared with the commercial vaccine group， the difference was not significant （P>0.05）. After challenged with PRV-AH， the protection rates were 100%， 70%， 60% and 90% of the rPRV-AH-gI-/gE-/gM-/gD+， rPRV-AH-gI-/gE-/gM-， rPRV-AH-gI-/gE- and commercial vaccine groups respectively. The results showed that the insertion of gD gene can improve the immune protection effect of the recombinant virus which is helpful in developing vaccine candidate for PRV control.
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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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