猪圆环病毒2型多抗原表位串联在大肠杆菌中表达与反应原性分析

Expression and identification of tandem multi-epitope of porcine circovirus type 2 in Escherichia coli

为将猪圆环病毒2型(PCV2)的多个抗原表位串联,并在大肠杆菌中表达,本研究人工合成PCV2多抗原表位串联的编码序列并与pET32a(+)载体连接构建重组表达质粒。将其转化到BL21感受态细胞,经IPTG诱导表达。检测结果显示,多抗原表位串联基因在大肠杆菌中获得表达并以包涵体形式存在;表达的融合蛋白分子量约为27 ku。将包涵体溶解并经Ni-His柱纯化得到纯化的目的蛋白,western blot和ELISA结果显示,该表达产物具有较好的反应原性。

To prepare efficient subunit vaccines against porcine circovirus type 2 （PCV2）, a sequence encoding multi-epitope of PCV2 was artificially synthesized and ligated into pET32a（＋） plasmid. The resultant plasmid was transformed into E. coli BL21 in which a fusion protein was expressed with IPTG induction. The SDS-PAGE analysis showed that fusion protein was approximately 27 ku, mainly in form of inclusion body. Western blot and ELISA detection indicated that the fusion protein reacted with positive PCV2 antibody. This study provided an alternative way to develop a potential PCV2 subunit vaccine and diagnostic reagents.