兔出血症病毒VP60主要抗原表位的原核表达及其免疫原性

Prokaryotic Expression and Immunogenicity of Major Antigen Epitope of Rabbit Haemorrhagic Disease Virus VP60

[目的]研究兔出血症病毒(RHDV)VP60主要抗原表位基因原核表达蛋白的免疫原性。[方法]采用RT-PCR方法扩增了RHDV衣壳蛋白VP60主要抗原表位基因。以pET-28b(+)为表达载体,在E.coliRosetta菌株中表达重组蛋白。通过Western blot分析和接种家兔检测了重组蛋白的免疫原性。[结果]经Western blot检测,在相对分子量24.0k Da处出现特异性的反应带。以纯化的重组蛋白制备的抗血清可以与纯化的RHDV发生特异性的ELISA反应。[结论]RHDV VP60主要抗原表位的原核表达产物具有较好的免疫原性。

[Objective]The paper aims to investigate the immunogenicity of main antigen epitope of RHDV （Rabbit haemorrhagic disease virus） VP60 which was expressed in a prokaryotic system.[Method]The major antigen epitope gene of RHDV VP60 was amplified by RT-PCR.It was cloned into pET-28b （＋ ） and expressed in E.coli Rosetta strain.The recombinant protein was detected by Western blot.The purified recombinant protein was used to immunize rabbits in order to observe its immunogenicity.[Result]Western blot analysis revealed a clear band at approximately 24.0 kDa.The purified recombinant protein reacted with the purified RHDV in ELISA.[Conclusion]The prokaryotically expressed main antigen epitope of RHDV VP60 shows good immunogenicity.