- 更多网络例句与包上外壳相关的网络例句 [注:此内容来源于网络,仅供参考]
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The minor core protein s C-encoding gene of Muscovy duck reovirus was cloned into theprokaryotic expression vector pET32a. The recombinant plasmid pET32a-s C was amplified andextracted after being transformed into E.coli DH5a competent cells. Restriction analysis withEcoRⅠand SacⅠand sequences analysis indicated that the recombinant plasmid was inserted withcorrect open reading frame. The fusion protein about 50 ku was produced after induction with 0.15mmol/L IPTG of E.coli competent cells transformed with pET32a-sC. The SDS-PAGE andWestern-bloting test indicated that the fusion protein reacted with the convalescents sera of duckinfected with Muscovy duck reovirus. The indirect ELISA method was developed by using thepurified fusion s C protein as coating antigen. The optimal concentration of s C was 5μg/ml, thedilution of serum sample was 1:40; The results showed that preparation of an ELISA by using sCas coating antigen in detecting 50 field duck sera in comparison with the AGIP were more sensitiveand specific than agar gel immuno-diffusion AGIP test. The results suggest that presence ofantibody against viral protein sC in duck may be a good indicator by the sC-ELISA for detectionof duck infection with reovirus.
同时,本研究将编码外壳蛋白σC的基因克隆于原核表达载体pET32a上,经过EcoRⅠ和SacⅠ双酶切鉴定和序列分析后,得到阳性重组质粒pET32a-σC;将阳性重组质粒pET32a-σC转化到大肠杆菌BL-21感受态细胞中进行诱导表达,经SDS-PAGE和Western-blbtting检测分析,融合表达的蛋白能够与番鸭呼肠孤病毒感染的康复鸭血清发生特异性反应;将融合表达的蛋白纯化后作为包被抗原,建立了检测鸭血清中呼肠孤病毒抗体的间接酶联免疫吸附试验检测方法,此方法中抗原的最佳包被浓度为5μg/ml、标准阳性血清的最适稀释倍数为1:40倍,用此方法对50份鸭血清样品进行检测,并与琼脂糖凝胶扩散试验检测抗体的法相比较,证明此ELISA方法具有良好的特异性和敏感性,本研究为今后鸭呼肠孤病毒诊断试剂盒的研制奠定了基础。
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