A DNA replicon system for rapid high-level production of virus-like particles in plants.
A DNA replicon system for rapid high-level production of virus-like particles in plants.
复制标题
DOI：
10.1002/bit.22299
复制
发表时间：
2009-07-01
影响因子：
3.8
通讯作者：
Mason, Hugh
中科院分区：
文献类型：
--
作者：
Huang, Zhong;Chen, Qiang;Hjelm, Brooke;Arntzen, Charles;Mason, Hugh
重组病毒样颗粒（VLP）代表了一种安全有效的疫苗策略。我们先前描述了一种稳定的转基因植物系统，用于廉价生产和口服递送VLP疫苗。然而，相对低水平的抗原积累和生产转基因植物的较长时间框架是基于植物的VLP生产的实际发展中的两个主要障碍。在本文中，我们描述了双生病毒衍生的DNA复制子载体的快速，高产的植物为基础的VLP生产的优化。通过农杆菌渗入本氏烟草叶共递送豆黄矮病毒（BeYDV）衍生的载体和Rep/RepA供应载体导致有效的复制子扩增和在五天内的稳健蛋白质生产。共表达番茄丛矮病毒P19蛋白，基因沉默抑制剂，通过稳定mRNA进一步增强VLP积累。利用该系统，分别以0.80和0.34 mg/g叶鲜重产生了B型肝炎核心抗原（HBc）和诺瓦克病毒衣壳蛋白（NVCP）。瞬时表达抗原的沉降分析和电子显微镜证实了VLP的有效组装。此外，一个单一的复制子载体含有一个内置的Rep/RepA盒没有p19驱动蛋白质表达在类似的水平作为三组分系统。这些结果证明了使用BeYDV衍生的DNA复制子系统在植物中瞬时表达快速和高水平生产基于VLP的疫苗的优点。
Recombinant virus-like particles (VLPs) represent a safe and effective vaccine strategy. We previously described a stable transgenic plant system for inexpensive production and oral delivery of VLP vaccines. However, the relatively low level antigen accumulation and long time frame to produce transgenic plants are the two major roadblocks in the practical development of plant-based VLP production. In this paper, we describe the optimization of geminivirus-derived DNA replicon vectors for rapid, high-yield plant-based production of VLPs. Co-delivery of bean yellow dwarf virus (BeYDV)-derived vector and Rep/RepA-supplying vector by agroinfiltration of Nicotiana benthamiana leaves resulted in efficient replicon amplification and robust protein production within five days. Co-expression of the P19 protein of tomato bush stunt virus, a gene silencing inhibitor, further enhanced VLP accumulation by stabilizing the mRNA. With this system, hepatitis B core antigen (HBc) and Norwalk virus capsid protein (NVCP) were produced at 0.80 and 0.34 mg/g leaf fresh weight, respectively. Sedimentation analysis and electron microscopy of transiently expressed antigens verified the efficient assembly of VLPs. Furthermore, a single replicon vector containing a built-in Rep/RepA cassette without p19 drove protein expression at similar levels as the three-component system. These results demonstrate the advantages of fast and high-level production of VLP-based vaccines using the BeYDV-derived DNA replicon system for transient expression in plants.
登录
查看更多内容
影响因子：
4
作者：
Ca単izares, MC;Nicholson, L;Lomonossoff, GP
通讯作者：
Lomonossoff, GP
影响因子：
56.9
作者：
HAQ, TA;MASON, HS;ARNTZEN, CJ
通讯作者：
ARNTZEN, CJ
影响因子：
5.5
作者：
Huang, Z;Elkin, G;Mason, HS
通讯作者：
Mason, HS
DOI：
10.2174/1568005014605973
发表时间：
2001-11-01
期刊：
Current Drug Targets - Infectious Disorders
影响因子：
--
作者：
Beyer, Thomas;Herrmann, Martin;Hess, Juergen
通讯作者：
Hess, Juergen
影响因子：
13.8
作者：
Huang, Zhong;LePore, Kate;Mason, Hugh S.
通讯作者：
Mason, Hugh S.