Expression of LamB Vaccine Antigen inWolffia globosa(Duck Weed) Against Fish Vibriosis
Expression of LamB Vaccine Antigen inWolffia globosa(Duck Weed) Against Fish Vibriosis
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DOI：
10.3389/fimmu.2020.01857
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发表时间：
2020-08-20
影响因子：
7.3
通讯作者：
Hou, Hongwei
中科院分区：
文献类型：
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作者：
Heenatigala, P. P. M.;Sun, Zuoliang;Hou, Hongwei
弧菌病是在咸水养殖的鱼类和贝类中发现的一种常见的细菌疾病。许多弧菌已被确定为病原体。Lamb是弧菌外膜蛋白(OMPS)家族的一员，在所有弧菌中都是保守的，被认为是预防弧菌病的有效候选疫苗。无根浮萍(Wolffia)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Wolffia中表达一种保护性的鱼弧菌疫苗抗原。我们扩增了溶藻弧菌的LamB基因，并对其进行了修饰以最大限度地提高蛋白的表达水平，并将蛋白转移到植物的内质网(ER)。将其克隆到CaMV 35S启动子控制的双元载体pMYC中，并将其导入Wolffia lobosaby农杆菌介导型转化。基因组聚合酶链式反应和逆转录-聚合酶链式反应证实了LamB基因的整合和表达。Western印迹分析表明，8个转基因品系中均有羊肉蛋白的积累。通过饲喂新鲜的转基因狼，然后用毒力V攻击斑马鱼，评价了转基因狼的交叉保护性能。溶藻胶菌。免疫鱼的高相对存活率(RPS)(63.3%)证实了转基因Wolffia免疫的鱼对弧菌感染有良好的保护作用。这些结果表明，Wolffia表达的羔羊可以作为鱼类弧菌病的可食用植物候选疫苗模型，以及利用Wolffiaas生物反应器生产可食用疫苗的可行性。
Vibriosis is a commonly found bacterial disease identified among fish and shellfish cultured in saline waters. A multitude ofVibriospecies have been identified as the causative agents. LamB, a member of outer membrane protein (OMPs) family of these bacteria is conserved among allVibriospecies and has been identified as an efficient vaccine candidate against vibriosis. Rootless duckweed (Wolffia) 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-transformedWolffia. We amplifiedLamBgene from virulentVibrio alginolyticusand 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 intoWolffia globosabyAgrobacterium-mediated transformation. Integration and expression of theLamBgene 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 transgenicWolffiawas evaluated by orally vaccinating zebrafish through feeding fresh transgenicWolffiaand subsequently challenging with virulentV. alginolyticus. High relative percent survival (RPS) of the vaccinated fish (63.3%) confirmed that fish immunized with transgenicWolffiawere well-protected fromVibrioinfection. These findings suggest thatWolffiaexpressed LamB could serve as an edible plant-based candidate vaccine model for fish vibriosis and feasibility of utilizingWolffiaas bioreactor to produce edible vaccines.