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目的:克隆并表达弓形虫SAG1基因成熟蛋白编码区片段,为下一步表达蛋白纯化奠定基础。 方法:将弓形虫SAG1基因成熟蛋白编码区片段克隆入高效融合表达载体pET32α,转化大肠杆菌BL21(DE3), 37℃、IPTG诱导表达,SDS PAGE分析表达产物。结果:成功构建重组质粒pET32α SAG1,经诱导后表达出含 外源基因的融合蛋白,SDS PAGE分析表达产物分子质量约44kDa。结论:弓形虫SAG1成熟蛋白编码区在原 核表达系统pET32α/BL21(DE3)中获得成功表达,为下一步表达蛋白纯化提供试验依据。
OBJECTIVE: To clone and express the mature protein coding region of SAG1 gene of Toxoplasma gondii, which will lay the foundation for further purification of the expressed protein. Methods: The fragment encoding the mature protein of SAG1 gene of Toxoplasma gondii was cloned into the highly efficient fusion expression vector pET32α, transformed into E. coli BL21 (DE3), induced by IPTG at 37 ℃, and expressed by SDS PAGE. Results: The recombinant plasmid pET32α SAG1 was successfully constructed and the fusion protein containing exogenous gene was expressed. The molecular weight of the expressed product was about 44 kDa by SDS PAGE. CONCLUSION: Toxoplasma gondii SAG1 mature protein coding region was successfully expressed in prokaryotic expression system pET32α / BL21 (DE3), which provided the experimental basis for the further purification of expressed protein.