Zhang G Gary, Rodrigues Lauren, Rovinski Benjamin, White K Andrew
Department of Biology, York University, Toronto, Ontario, Canada.
Mol Biotechnol. 2002 Feb;20(2):131-6. doi: 10.1385/MB:20:2:131.
The production of antigens for vaccines in plants has the potential as a safe and cost-effective alternative to traditional production systems. Toward the development of a plant-based expression system for the production of human immunodeficiency virus type I (HIV-1) p24 capsid protein, the p24 gene was introduced into the genome of tobacco plants using Agrobacterium tumefaciens-mediated gene transfer. Southern blot analyses confirmed the presence of the p24 coding sequence within the genome of transgenic lines. Western blot analysis of protein extracts from transgenic plants identified plant-expressed p24 protein that cross-reacted with a p24-specific monoclonal antibody, thus confirming the maintenance of antigenicity. Quantification of the p24 protein using enzyme-linked immunosorbent assay (ELISA) estimated yields of approx 3.5 mg per g of soluble leaf protein. Similar accumulation levels of p24 were also detected in T1 plants, confirming that the p24 gene is transmitted stably. Our results indicate that plant-based transgenic expression represents a viable means of producing p24 for the development of HIV vaccine and for use in HIV diagnostic procedures.
利用植物生产疫苗抗原，有望成为一种安全且具成本效益的传统生产系统替代方案。为开发用于生产人类免疫缺陷病毒I型（HIV-1）p24衣壳蛋白的植物表达系统，采用根癌农杆菌介导的基因转移方法，将p24基因导入烟草植物基因组。Southern杂交分析证实转基因株系基因组中存在p24编码序列。对转基因植物蛋白提取物进行Western杂交分析，鉴定出与p24特异性单克隆抗体发生交叉反应的植物表达p24蛋白，从而证实了抗原性的保持。使用酶联免疫吸附测定（ELISA）对p24蛋白进行定量分析，估计每克可溶性叶蛋白产量约为3.5毫克。在T1代植株中也检测到类似的p24积累水平，证实p24基因能够稳定遗传。我们的结果表明，基于植物的转基因表达是生产用于HIV疫苗开发及HIV诊断程序的p24的可行方法。
