【摘 要】
：
目的表达和纯化SARS冠状病毒的N蛋白,分析其诱导的免疫应答.方法从重组克隆pMD18-N中切取N基因的插入片段,亚克隆至原核表达质粒pET-23a(+)中,转化大肠杆菌BL21,PCR和酶切鉴
论文部分内容阅读
目的表达和纯化SARS冠状病毒的N蛋白,分析其诱导的免疫应答.方法从重组克隆pMD18-N中切取N基因的插入片段,亚克隆至原核表达质粒pET-23a(+)中,转化大肠杆菌BL21,PCR和酶切鉴定转化菌落的插入序列;将构建的原核表达菌株经IPTG诱导,SDS-PAGE分析重组蛋白的表达;大量诱导表达N蛋白,金属螯合层析予以纯化,将纯化蛋白免疫小鼠,观察其诱导的免疫应答.结果 N基因的特异片段被亚克隆到原核表达质粒pET-23a(+),在大肠杆菌表达了SARS冠状病毒的N蛋白;表达的蛋白经金属螯合层析获得纯化;纯化蛋白能被重组N蛋白的GST融合蛋白的免疫血清识别,并诱导小鼠产生了高滴度的抗体和Th1/Th2型的免疫应答.结论成功构建了N蛋白的原核重组表达质粒,纯化的蛋白具有良好的抗原性.
其他文献
Calpains are a group of calcium-dependent proteases that are over activated by increased intracellular calcium levels under pathological conditions. A wide rang
目的构建包含FMDV结构蛋白P1、非结构蛋白2A、3C以及部分2B的基因片段P12X3C的植物组成型表达载体,为FMDV可饲疫苗的研制奠定基础.方法通过PCR扩增法,从pGEM/P12A质粒和pGEM/
目的研究慢性应激对强迫游泳大鼠海马神经元热休克蛋白70(Hsp70)表达的影响.方法将50只大鼠随机分为实验组和对照组各25只.实验组大鼠通过21天的应激刺激制作抑郁动物模型,此
The neuroprotective effect against spinal cord ischemia/reperfusion injury in rats exerted by delayed xenon post-conditioning is stronger than that produced by
目的检测采自野外株和实验室驯化株白纹伊蚊对溴氰菊酯的抗性水平,探讨实验室抗性株(R-lab)的抗性机理.方法以采自深圳、广州龙洞和东莞庄的三株白纹伊蚊(SZ,LD和DW)和两株实
Rho kinase (ROCK) was the first downstream Rho effector found to mediate RhoA-induced actin cytoskeletal changes through effects on myosin light chain phosphory
Previous studies have reported age-specific pathological and functional outcomes in young and aged patients suffering spinal cord injury, but the mechanisms rem
Visual field defect is one of the various clinical manifestations in patients with subarachnoid hemorrhage (SAH). Little is known about the pathogenic mechanism
刚刺颚口线虫(Gnathostoma hispidum)是狗、猫、虎、豺等食肉动物的寄生虫,偶可感染人体〔1-2〕.成虫寄生于终宿主胃壁中,肿块破溃后虫卵入肠腔随粪便排出.迄今,我国已报告的