Liu Qing, Li Jin, Li Run-Li, Ma Ye-Ting, Zhou Peng-Lai, Gao Wen-Wei, Ma Hai-Li
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, 030801 Shanxi People's Republic of China.
3 Biotech. 2020 Mar;10(3):86. doi: 10.1007/s13205-020-2079-8. Epub 2020 Feb 4.
The development of rapid, simple, and sensitive diagnostic methods for identification of avian infectious bronchitis virus (IBV) is crucial for the effective control of avian infectious bronchitis. In the present study, a tandemly arranged multiepitope peptide (named SEMN) was designed with four antigenic regions derived from four major structural proteins of IBV. Then, we performed codon optimization of SEMN gene by changing the codon-adaptation index from 0.45 to 0.94 and expressed the optimized gene in codon bias-adjusted Rosetta (DE3), followed by determination of the immunoreactivity of the purified protein. Bioinformatics analysis of SEMN showed a high antigenicity, surface probability and hydrophilicity. The recombinant protein rSEMN was expressed both in soluble forms and as inclusion bodies, and the molecular weight of rSEMN was about 39 kDa. The preliminary diagnostic performance of rSEMN was confirmed by Western blotting analysis using chicken anti-IBV polyclonal antibodies. Further studies are needed to evaluate the immunogenicity in animal models and to give a final assessment of the diagnostic utility of this recombinant multi-epitope antigen.
开发快速、简单且灵敏的诊断方法以鉴定禽传染性支气管炎病毒（IBV）对于有效控制禽传染性支气管炎至关重要。在本研究中，设计了一种串联排列的多表位肽（命名为SEMN），其具有来自IBV四种主要结构蛋白的四个抗原区域。然后，我们通过将密码子适应指数从0.45改变为0.94对SEMN基因进行密码子优化，并在密码子偏性调整后的Rosetta（DE3）中表达优化后的基因，随后测定纯化蛋白的免疫反应性。对SEMN的生物信息学分析显示出高抗原性、表面概率和亲水性。重组蛋白rSEMN以可溶性形式和包涵体形式表达，rSEMN的分子量约为39 kDa。使用鸡抗IBV多克隆抗体通过蛋白质印迹分析证实了rSEMN的初步诊断性能。需要进一步研究以评估其在动物模型中的免疫原性，并对这种重组多表位抗原的诊断效用进行最终评估。
