摘要
目的表达和纯化弓形虫表面抗原1(SAG1),并分析其免疫学活性。方法构建pET15b-SAG1质粒,转化到宿主菌E.coil BL21(DE3)中,经IPTG诱导表达,使用Ni 2+亲和层析柱对重组蛋白进行纯化,用间接ELISA检测目的蛋白的抗原性。结果 SDS-PAGE显示表达的SAG1蛋白为可溶性的,经纯化后其纯度可达到90%以上;ELISA检测表达蛋白能被弓形虫感染者血清识别;棋盘滴定显示重组SAG1蛋白适宜包被浓度为0.5μg/ml。结论本研究表达的SAG1蛋白为可溶性的,且具有良好的抗原性,为进一步建立弓形虫血清学诊断方法和研制弓形虫病疫苗奠定了基础。
Objective To clone and express Toxoplasma gondii surface antigen1（SAG 1） and to analyze the immunological activity of the expressed protein SAG1. Methods The recombinant plasmid pET15b-SAG1 was constructed and then was transformed to Escherichia coil BL21（DE3） for expression under induction of IPTG.The recombinant protein was purified by metal-chelating affinity chromatography using Ni2＋.ELISA was used to analyze the antigenicity of the recombinant antigen protein.Results SDS-PAGE indicated that the fusion protein of SAG1 was expressed in supernatant at a purity of more than 90%.ELISA results indicated that the recombinant protein was recognized by toxoplasmosis-positive serum with a 0.5 μg/ml working concentration.Conclusion The SAG1 protein expressed in this experiment was soluble and had good antigenicity.These findings have laid the foundation for detection of Toxoplasma antibody or development of a Toxoplasma vaccine.
出处
《中国病原生物学杂志》
CSCD
2011年第11期816-818,812,共4页
Journal of Pathogen Biology
关键词
弓形虫
SAG1蛋白
蛋白表达
纯化
抗原性
Toxoplasma
SAG1 protein
protein expression
protein purification
antigenicity
