Conditional screening of recombinant proteins of Dermacentor marginatus AQPs and anti-tick effect
10.16303/j.cnki.1005-4545.2024.07.16
- VernacularTitle:边缘革蜱AQPs重组蛋白的表达及抗蜱效果
- Author:
Jun WU
1
;
Wenwen HE
;
Hao PU
;
Min JIN
;
Wenyu SHI
;
Aijun MA
;
Tingxiang LUO
;
Depeng YANG
;
Bayinchahan
;
Ercha HU
Author Information
1. 新疆农业大学动物医学学院,新疆乌鲁木齐 830052
- Keywords:
Dermacentor marginatus;
aquaporin;
preparation of polyclonal antibodies;
evaluation of anti-tick effect
- From:
Chinese Journal of Veterinary Science
2024;44(7):1466-1472,1506
- CountryChina
- Language:Chinese
-
Abstract:
In order to mine candidate vaccine antigens against ticks and to control ticks safely and effectively,the aim of this study was to immunize rabbits with purified aquaporins(AQPs)rD-mAQP1,rDmAQP2 and rDmAQP3 of Dermacentor marginatus.Blood collections for Western blot and ELISA tests were performed.The anti-tick challenge was conducted.The optimal expression conditions of rDmAQP1,rDmAQP2 and rDmAQP3 were screened by SDS-PAGE gel electrophore-sis.The three recombinant proteins were purified by HisSepNi-NTA6FF purification column.Rab-bits were divided into four groups of three rabbits each,including a control group and three immu-nized groups.The three purified recombinant proteins were separately immunized to three groups of rabbits,and the rabbits were immunized once on the 0th,14th and 21st day.Blood samples were collected every 7 days to prepare polyclonal antibodies.The reactivity was detected by Western blot and the antibody titer was detected by ELISA.Tick challenge test was carried out after the anti-body titer increased.The results showed that the optimal expression conditions for rDmAQP1 were induced for 8 h at IPTG concentration of 1.0 mmol/L and 37 ℃;the optimal expression conditions for rDmAQP2 were induced for 7 h at IPTG concentration of 1.0 mmol/L and 37 ℃;and the opti-mal expression conditions for rDmAQP3 were induced for 5 h at IPTG concentration of 1.0 mmol/L and 37 ℃.Western blot results showed that rDmAQP1,rDmAQP2 and rDmAQP3 all had certain reactivity.The ELISA results showed that the antibody titers of rabbits immunized with rD-mAQP1,rDmAQP2 and rDmAQP3 were as follows:the total anti-tick effect of rDmAQP1 protein was 79.74％,and the inhibition rates on average full-blooded tick weight,average egg weight and average egg hatching rate were 9.43％,25.17％and 63.81％,respectively.The total anti-tick effect of rDmAQP2 protein was 78.78％,and the inhibition rates of average full-blooded tick weight,av-erage egg weight and average egg hatching rate of Dermacentor marginatus were 8.30％,20.14％and 68.26％,respectively.The total anti-tick effect of rDmAQP3 protein was 87.91％,and the inhi-bition rates of average full-blooded tick weight,average egg weight and average egg hatching rate were 3.23％,22.47％and 80.5％,respectively.Through serological test and anti-tick test,it has been found that rDmAQP1,rDmAQP2 and rDmAQP3 all have the potential of candidate antigens against ticks,among which rDmAQP3 has the best immune effect,which lays a foundation for the study of the function of rDmAQP1,rDmAQP2 and rDmAQP3.
