| 摘要: |
| 【目的】制备大口黑鲈蛙虹彩病毒主衣壳(MCP)蛋白兔多克隆抗体，以期为该病毒蛋白功能研究奠定基础。【方法】对pET32a(+)MCP/BL21重组大肠杆菌进行诱导表达，将重组蛋白进行纯化、复性，以此为抗原免疫大耳兔制备多克隆抗体，ELISA检测抗体效价，间接免疫荧光试验（IFA）和Western Blot 法分析MCP多克隆抗体的特异性。【结果】纯化的MCP重组蛋白条带特异；间接ELISA结果显示，制备的兔多克隆抗体血清效价为1∶1 024 000；IFA和Western Blot检测结果表明该多抗特异性良好，能够与大口黑鲈蛙虹彩病毒MCP蛋白发生特异性反应，IFA试验表明血清最适稀释度为1∶500。【结论】成功制备了大口黑鲈蛙虹彩病毒MCP兔多克隆抗体，该抗体可特异性识别大口黑鲈蛙虹彩病毒MCP蛋白。 |
| 关键词: 大口黑鲈 蛙虹彩病毒 主衣壳蛋白 多克隆抗体 |
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| 基金项目:广东省海洋经济发展（海洋六大产业）专项资金项目(GDOE［2022］A13)；中国水产科学研究院中央级公益性科研院所基本科研业务费专项（2020TD45）；广东省现代农业产业技术创新团队专项基金项目(2019KJ140,2019KJ141) |
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| Preparation and application of polyclonal antibody against MCP protein of largemouth bass ranavirus |
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WANG Ruoxuan,UO Xia,LI Ningqiu,et al
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| Abstract: |
| 【Objective】The rabbit polyclonal antibody against recombinant MCP protein of largemouth bass ranavirus (LMBV) were prepared to provide basis for investigating LMBV protein functions.【Method】The pET32a(+)MCP/BL21 recombinant E.coli was induced to express,and the recombinant protein was purified and renatured.The polyclonal antibody was prepared by immunizing rabbits with the recombinant protein.The antibody titer was detected by ELISA and the specificity of MCP polyclonal antibody was analyzed by indirect immunofluorescence assay (IFA) and Western Blot.【Result】The purified MCP recombinant protein was specific.Indirect ELISA results showed that the serum titer of rabbit polyclonal antibody was 1∶1 024 000.The results of IFA and Western Blot illustrated polyclonal antibody had good specificity to LMBV.The optimum dilution of serum in IFA test was 1∶500.【Conclusion】The polyclonal antibody against MCP of LMBV was successfully prepared with specific recognition of major capsid proteins of largemouth bass ranavirus. |
| Key words: Micropterus salmoides ranavirus major capsid protein polyclonal antibody |
