Institute of Experimental Botany, v. v. i., Academy of Sciences of Czech Republic, Na Karlovce 1a, 16000 Prague 6, Czech Republic.
J Biosci. 2012 Mar;37(1):125-33. doi: 10.1007/s12038-011-9177-z.
Transient expression of foreign genes based on plant viral vectors is a suitable system for the production of relevant immunogens that can be used for the development of a new generation of vaccines against a variety of infectious diseases. In the present study the epitope derived from HPV-16 L2 minor capsid protein (amino acids 108-120) was expressed from Potato virus X (PVX)-based vector pGR106 as N- or C-terminal fusion with the PVX coat protein (PVX CP) in transgenic Nicotiana benthamiana plants. The fusion protein L2 108-120-PVX CP was successfully expressed in plants at a level of 170 mg/kg of fresh leaf tissue. The C-terminal fusion protein PVX CP- L2 108-120 was expressed using mutated vector sequence to avoid homologous recombination at a level of 8 mg/kg of fresh leaf tissue. Immunogenicity of L2 108-120-PVX CP virus-like particles was tested after immunization of mice by subcutaneous injection or tattoo administration. In animal sera the antibodies against the PVX CP and the L2 108-120 epitope were found after both methods of vaccine delivery.
基于植物病毒载体的外源基因瞬时表达是生产相关免疫原的合适系统，可用于开发针对多种传染病的新一代疫苗。本研究将 HPV-16 L2 次要衣壳蛋白（氨基酸 108-120）的表位从 Potato virus X（PVX）为基础的载体 pGR106 表达为 N-或 C-末端与 PVX 外壳蛋白（PVX CP）融合在转基因 Nicotiana benthamiana 植物中。融合蛋白 L2 108-120-PVX CP 在植物中成功表达，新鲜叶片组织的含量为 170mg/kg。使用突变载体序列在新鲜叶片组织的 8mg/kg 水平上表达 C-末端融合蛋白 PVX CP-L2 108-120，以避免同源重组。通过皮下注射或纹身给药对小鼠进行免疫接种后，测试了 L2 108-120-PVX CP 病毒样颗粒的免疫原性。在两种疫苗接种方法后，在动物血清中均发现了针对 PVX CP 和 L2 108-120 表位的抗体。
