Varsani Arvind, Williamson Anna-Lise, Stewart Debbie, Rybicki Edward P
Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, Cape Town 7701, South Africa.
Virus Res. 2006 Sep;120(1-2):91-6. doi: 10.1016/j.virusres.2006.01.022. Epub 2006 Mar 13.
A Tobacco mosaic virus (TMV)-derived vector was used to express a native Human papillomavirus type 16 (HPV-16) L1 gene in Nicotiana benthamiana by means of infectious in vitro RNA transcripts inoculated onto N. benthamiana plants. HPV-16 L1 protein expression was quantitated by enzyme-linked immunosorbent assays (ELISA) after concentration of the plant extract. We estimated that the L1 product yield was 20-37 microg/kg of fresh leaf material. The L1 protein in the concentrated extract was antigenically characterised using the neutralising and conformation-specific Mabs H16:V5 and H16:E70, which bound to the plant-produced protein. Particles observed by transmission electron microscopy were mainly capsomers but virus-like particles (VLPs) similar to those produced in other systems were also present. Immunisation of rabbits with the concentrated plant extract induced a weak immune response. This is the first report of the successful expression of an HPV L1 gene in plants using a plant virus vector.
利用烟草花叶病毒（TMV）衍生载体，通过将体外转录的感染性RNA接种到本氏烟草植株上，在本氏烟草中表达天然人乳头瘤病毒16型（HPV - 16）L1基因。在浓缩植物提取物后，通过酶联免疫吸附测定（ELISA）对HPV - 16 L1蛋白表达进行定量。我们估计L1产物产量为每千克鲜叶材料20 - 37微克。使用中和性和构象特异性单克隆抗体H16:V5和H16:E70对浓缩提取物中的L1蛋白进行抗原特性分析，这两种抗体可与植物产生的蛋白结合。透射电子显微镜观察到的颗粒主要是衣壳粒，但也存在类似于其他系统产生的病毒样颗粒（VLP）。用浓缩植物提取物免疫兔子诱导出较弱的免疫反应。这是首次报道利用植物病毒载体在植物中成功表达HPV L1基因。
