摘要
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 gene coding mature peptide of SAG1 of Toxoplasma gondii for its pure protein studies. Methods:The gene region coding mature peptide of SAG1 was amplified by PCR and inserted into expression vector pET32α,then recombinant plasmid pET32α-SAG1 was transformed into E.coli BL21(DE3).The transformants pET32α-SAG1 were induced by IPTG at the temperature of 37 ℃ and the expressed protein was analyzed by SDS-PAGE. Results:The recombinant plasmid was constructed successfully.SDS-PAGE analysis of bacterial crude extracts showed that the molecular weight of the expressed fused proteins was about 44 kDa. Conclusion:SAG1 mature peptide coding gene can be expressed in E.coli and the successfully expressed protein could be the foundation of further studies of SAG1.
出处
《广州医学院学报》
2004年第4期14-16,共3页
Academic Journal of Guangzhou Medical College
基金
广州医学院自然科学基金(编号:01-K-33)
