原核表达番鸭呼肠孤病毒σC与σB蛋白对雏番鸭的免疫保护

doi:10.3864/j.issn.0578-1752.2013.14.020

摘要/Abstract

摘要： 【目的】探讨原核表达的番鸭呼肠孤病毒（DRV）外壳蛋白σC与σB对于雏番鸭的免疫保护作用，为研制有效的DRV亚单位疫苗提供理论基础。【方法】对pET-30a-S3/BL21重组菌与pET-30a-S4 ORF2/BL21重组菌进行稳定性检验，表达的重组蛋白进行纯化和复性后，进行油乳剂制备，50只1日龄雏番鸭随机平均分为5组，即DRV σB蛋白免疫组、DRV σC蛋白免疫组、DRV σB + σC蛋白免疫组及PBS对照组和攻毒对照组，免疫组皮下接种剂量为500 μg/只，7日龄时进行二次免疫，14日龄时进行攻毒试验，ELISA和琼扩（AGP）检测抗体水平并计算保护指数。【结果】重组菌具有良好的稳定性，各免疫组的雏番鸭均在第2次免疫后7 d产生了一定效价的抗体，其中DRV σB + σC蛋白免疫组和DRV σC蛋白免疫组的雏番鸭血清抗体ELISA效价均为1﹕200，高于DRV σB蛋白免疫组的1﹕100。攻毒保护试验结果表明，联合免疫的保护指数最高，可达70，重组DRV σC保护指数次之，为60，重组DRV σB蛋白保护指数最低，仅为40。【结论】使用原核表达的DRV外壳蛋白σC与σB制备油乳剂，联合免疫可诱导雏番鸭快速产生一定效价的抗体，并提供良好的免疫保护力。

关键词: 番鸭呼肠孤病毒 , &sigma, C与&sigma, B蛋白 , 免疫保护

Abstract: 【Objective】 The immunogenicity of the recombinant Musovy Duck Reovirus (DRV) capsid proteins (σC and σB) expressed in E. coli was investigated and thus the base of the research of effective subunit vaccine for DRV was established. 【Method】The stability of the recombinant E.coli pET-30a-S3/BL21 and pET-30a-S4 ORF2/BL21 were detected, while the recombinant proteins were emulsified after purification and renaturation. Fifty young musovy ducks were randomly divided into 5 groups including 3 treatment groups, the PBS control group and the infection control group. The treatment groups were injected subcutaneously with a dosage of 500 μg per injection, on 1 d and 7 d respectively. The recombinant DRV σB protein, the recombinant DRV σC protein, as well as both σB and σC proteins were used. On the 14d, the animals except the PBS control group, were challenged with YB isolate of DRV. The titer of the antibody raised by the recombinant proteins and the protection rate were detected. 【Result】The recombination E.coli was stable. The titers of the antibody of the immunity groups were all raised on the 7th day after the second injection. It was found that the titer of antibody in the group injected with both σB and σC proteins as well as the group injected with recombinant σC was 1﹕200, but in the meanwhile, that in the group injected with recombinant σB protein was just 1﹕100. The protection test showed that the injection with both σB and σC proteins could provide best protection for DRV infection with protection index 70, and then the second was the group injected with DRV σC protein with 60, while the last was the group with DRV σB protein with 40. 【Conclusion】The injection with both recombinant σB and σC protein expressed in E.coli and emulsified could provide protection to young musovy ducks resisting DRV infection.

Key words: Musovy Duck Reovirus , σC and σB protein , immunization

吴晓平, 吴异健, 韩典霖, 吴宝成, 黄一帆. 原核表达番鸭呼肠孤病毒σC与σB蛋白对雏番鸭的免疫保护[J]. 中国农业科学, 2013, 46(14): 3040-3045.

WU Xiao-Ping, WU Yi-Jian, HAN Dian-Lin, WU Bao-Cheng, HUANG Yi-Fan. Protection of Musovy Duck Reovirus by Immunization with the Recombinant σC and σB Protein Expressed in E. coli[J]. Scientia Agricultura Sinica, 2013, 46(14): 3040-3045.

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