University of Chinese Academy of Sciences, Beijing, China.
The State Key Laboratory of Freshwater Ecology and Biotechnology, The Key Laboratory of Aquatic Biodiversity and Conservation of Chinese Academy of Sciences, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China.
Front Immunol. 2020 Aug 20;11:1857. doi: 10.3389/fimmu.2020.01857. eCollection 2020.
Vibriosis is a commonly found bacterial disease identified among fish and shellfish cultured in saline waters. A multitude of species have been identified as the causative agents. LamB, a member of outer membrane protein (OMPs) family of these bacteria is conserved among all species and has been identified as an efficient vaccine candidate against vibriosis. Rootless duckweed () is a tiny, edible aquatic plant possessing characteristics suitable for the utilization as a bioreactor. Thus, we attempted to express a protective edible vaccine antigen against fish vibriosis in nuclear-transformed . We amplified gene from virulent and it was modified to maximize the protein expression level and translocate the protein to the endoplasmic reticulum (ER) in plants. It was cloned into binary vector pMYC under the control of CaMV 35S promoter and introduced into by -mediated transformation. Integration and expression of the gene was confirmed by genomic PCR and RT-PCR. Western blot analysis revealed accumulation of the LamB protein in 8 transgenic lines. The cross-protective property of transgenic was evaluated by orally vaccinating zebrafish through feeding fresh transgenic and subsequently challenging with virulent . High relative percent survival (RPS) of the vaccinated fish (63.3%) confirmed that fish immunized with transgenic were well-protected from infection. These findings suggest that expressed LamB could serve as an edible plant-based candidate vaccine model for fish vibriosis and feasibility of utilizing as bioreactor to produce edible vaccines.
弧菌病是一种常见的细菌性疾病,在盐水中养殖的鱼类和贝类中发现。已经鉴定出多种物种是病原体。LamB 是这些细菌的外膜蛋白 (OMPs) 家族的成员,在所有物种中都保守存在,已被鉴定为针对弧菌病的有效疫苗候选物。无根浮萍()是一种微小的、可食用的水生植物,具有适合用作生物反应器的特性。因此,我们试图在核转化的中表达一种针对鱼类弧菌病的保护性可食用疫苗抗原。我们从毒力中扩增了基因,并对其进行了修饰,以最大限度地提高蛋白质表达水平并将蛋白质易位到植物内质网 (ER) 中。它被克隆到 pMYC 双元载体中,受 CaMV 35S 启动子的控制,并通过介导转化引入。通过基因组 PCR 和 RT-PCR 证实了的整合和表达。Western blot 分析显示 8 个转基因系中积累了 LamB 蛋白。通过口服喂食新鲜转基因来评估转基因的交叉保护特性通过喂食新鲜转基因来评估转基因的交叉保护特性并随后用毒力挑战。接种鱼的相对百分比存活(RPS)较高(63.3%)证实,用转基因免疫的鱼可以很好地免受感染。这些发现表明,表达的 LamB 可以作为鱼类弧菌病的可食用植物疫苗候选物模型,并且利用作为生物反应器生产可食用疫苗是可行的。