非典型性肺炎S蛋白抗原决定簇序列的串联合成和表达

Synthesizing and expressing antigenic determinant sequence of S protein of SARS

非典型性肺炎病毒的S蛋白是病毒表面最主要的膜蛋白,它在病毒与受体结合和病毒的早期感染过程中起重要的作用。S蛋白也是SARS病毒主要的免疫原,是用来研制保护性疫苗的理想部位。研究表明S蛋白的抗原表位比较多且分散,并且由于分子量比较大,因此对其抗原表位的研究就有重要意义。根据已有的资料报道和软件分析,本实验将S蛋白的主要抗原表位及受体结合区串联在一起,利用基因合成的方法合成了这段串联基因。并通过基因操作构建了表达合成基因的原核重组质粒pET-28a-S合,转化大肠杆菌并诱导目的蛋白的表达,SDS-PAGE检测结果表明,重组菌能够表达目的蛋白,其表达量占菌体总蛋白的23.5%。Westernblot分析结果证实该重组蛋白具有良好的免疫原性。

S protein is the mainly membrane protein of SARS. It responses for binding to specific cellular receptors and plays an important role in the earlier infection. S protein is the mainly antigen of SARS, and could be a good candidate for further developing SARS vaccine. From the previous studies, the antigenic determinant sequence of S protein is separated and the molecular weight of S protein is too large, so it is important to study the antigenic determinant sequence of S protein. Accorded to the material and analysis by soft-ware, the antigenic determinant sequence and the receptors binding sequence was jointed together and synthesized. The synthesized gene was ligated to pET-28a and constructed the recombinant plasmid pET-28a-Ssynthesized. The recombinant vector was transformed into E.coli BL21 and induced with IPTG, the targeted protein was detected by SDS-PAGE. The Western blot experiment showed it was immunogenic.