摘要
[目的]探讨弓形虫微线体蛋白MIC3基因在大肠杆菌内高效表达的条件。[方法]通过SDS-PAGE分析,研究不同菌液密度、诱导剂IPTG浓度和诱导时间对MIC3基因在大肠杆菌中表达量的影响。[结果]MIC3基因在大肠杆菌中最适表达条件为:菌液密度OD600为0.4,诱导剂IPTG终浓度为0.4mmol/L和诱导时间3.5h。在该条件下表达的融合蛋白经初步分离提纯后,1 L培养液约含融合蛋白143mg,目的蛋白表达量占菌体总蛋白的49%。[结论]该研究为研究和制备弓形虫病的rMIC3诊断试剂奠定了基础。
[Objective] The research aimed to discuss the conditions of high-level expression of MIC3 gene of microneme protein of Toxoplasma gondii in Escherichia coli. [Method] Through SDS-PAGE analysis, the effects of different density of bacteria liquid, the concn, of inducer IPTG and induction time on the expression of MIC3 gene in E. coli. [Result] The optimum expression conditions of MIC3 gene in E. coli were as follows： the density OD600 of bacteria liquid was 0.4, the final concn, of inducer IPTG was 0.4 mmol/L and the induction time was 3.5 h. Under these conditions, there were 143 g fusion protein in 1 L culture liquid after the expressed fusion protein was preliminarily isolated and purified. The expression amount of target protein occupied 49 % of total bacteria protein. [Conclusion] This research laid the foundation for studying and preparing rMIC3 diagnostic reagent against the disease of Toxoplasmagondii.
出处
《安徽农业科学》
CAS
北大核心
2008年第17期7157-7158,共2页
Journal of Anhui Agricultural Sciences
关键词
弓形虫
MIC3基因
高效表达
Toxoplasma gondii
MIC3 gene
High-level expression
