| 摘要: |
| [目的]构建猪瘟病毒(Classical Swine Fever Virus,CSFV)E2基因主要抗原区原核表达质粒载体,高效表达E2蛋白,为进一步研究E2亚单位疫苗提供物质基础.[方法]用RT-nested PCR技术扩增CSFV流行毒株SXYL株E2基因主要抗原区,克隆于表达载体pET32a中,成功构建表达质粒pET32a-E2,将pET32a-E2转入BL21(DE3)受体菌并用IPTG诱导表达E2蛋白,对E2蛋白进行SDS-PAGE电泳、Western-blot分析和可溶性鉴定.[结果]CSFV E2基因得到了高效表达,表达蛋白量占菌体蛋白量的33.2%,表达蛋白E2主要以包涵体形式存在;免疫印迹分析结果表明,表达蛋白E2能识别天然猪瘟多克隆抗体.[结论]E2蛋白表达量高并具有生物学活性. |
| 关键词: 猪瘟 E2基因 原核表达 |
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| Study on pronucleus expression of key antigen region domain E2 gene of Classical Swine Fever Virus prevailing strain in E. coli |
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| Abstract: |
| 【Objective】 The study is to construct plasmid vector of key antigen region domain CSFV E2 gene and express E2 protein highly efficient,to offer substantial foundation for studding E2 subunit vaccine.【Method】 Clonied key antigen region domain E2 gene of strain SXYL by RT-nested PCR,connected it with pET32a to fabricate expressing plasmid pET32a-E2 successfully,translated pET32a-E2 to BL21（DE3）and induced it express E2 protein by IPTG,identified E2 protein by SDS-PAGE,Western-blot and ultrasonic.【Result】 Recombined E.coli could express E2 protein highly efficient,the quantity of expression is 33.2％,most E2 protein is inclusion bodies,the E2 protein could identify natural polyclonal antibody of CSFV.【Conclusion】 The quantity of expression protein is high and it has biological activity. |
| Key words: Classical Swine Fever E2 gene pronucleus expression |
