Abstract：
Transient expression is the major method to express foot-and-mouth disease virus (FMDV) capsid proteins in mammalian cells. To achieve stable expression of FMDV capsid proteins and efficient assembly of virus like particles (VLPs) in cells, the plasmids of piggyBac (PB) transposon-constitutive expression and PB transposon-tetracycline (Tet) inducible expression vectors were constructed. The function of the plasmids was tested by fluorescent proteins. By adding antibiotics, the constitutive cell pools (C-WT, C-L127P) expressing P12A3C (WT/L127P) genes and the inducible cell pools (I-WT, I-L127P) expressing P12A3C (WT/L127P) genes were generated. The genes of green fluorescent protein, 3C protease and reverse tetracycline transactivator (rtTA) were integrated into chromosome, which was confirmed by fluorescence observation and PCR testing. The cell pool I-L127P has a stronger production capacity of capsid proteins and VLPs, which was confirmed by Western blotting and enzyme linked immunosorbent assay (ELISA), respectively. In conclusion, inducing the chromosomal expression of FMDV capsid proteins was firstly reported, which may facilitate the technical process of mammalian production of FMDV VLPs vaccine and the construction of mammalian inducible expression systems for other proteins.
摘要：
瞬时表达是在哺乳动物细胞中表达口蹄疫病毒（FMDV）衣壳蛋白的主要方法。为了实现FMDV衣壳蛋白的稳定表达和病毒样颗粒(VLPs)在细胞中的高效组装,构建piggyBac（PB）转座子组成型表达质粒和PB转座子-四环素（Tet）诱导型表达载体。通过荧光蛋白测试质粒的功能。通过添加抗生素，组成型细胞池(C-WT，C-L127P）表达P12A3C（WT/L127P）基因和可诱导细胞池（I-WT，产生了表达P12A3C(WT/L127P)的I-L127P)基因。绿色荧光蛋白的基因，3C蛋白酶和反向四环素反式激活剂（rtTA）整合到染色体上，荧光观察和PCR检测证实了这一点。细胞池I-L127P具有更强的衣壳蛋白和VLP生产能力，通过蛋白质印迹和酶联免疫吸附测定（ELISA）证实，分别。总之,首次报道了诱导FMDV衣壳蛋白的染色体表达，这可能有助于哺乳动物生产FMDVVLP疫苗的技术过程以及其他蛋白质的哺乳动物诱导型表达系统的构建。
