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通过转基因油菜籽经口投喂方式向鸡传递新型嵌合血凝素-神经氨酸酶融合抗原的免疫原性研究。

The Immunogenicity of a Novel Chimeric Hemagglutinin-Neuraminidase-Fusion Antigen from Newcastle Disease Virus by Oral Delivery of Transgenic Canola Seeds to Chickens.

机构信息

Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Shahrake- Pajoohesh Blvd. 15th Km, Tehran-Karaj Highway, P.O. Box 14965-161, Tehran, Iran.

Agricultural Research Education and Extension Organization, Razi Vaccine and Serum Research Institute, Karaj, Iran.

出版信息

Mol Biotechnol. 2020 Jul;62(6-7):344-354. doi: 10.1007/s12033-020-00254-y.

Abstract

Newcastle disease (ND) is considered as one of the most devastating infectious diseases targeting domestic birds and has considerable threat to the commercial poultry production. Two surface glycoproteins, hemagglutinin-neuraminidase (HN) and fusion (F), act as antigens in the virus structure and also play important roles in infecting host cells. In the current study, the expression of the chimeric HN-F protein in canola seeds and its immunogenicity in chickens were investigated. The HN-F gene was cloned downstream of the fatty acid elongase 1 (FAE1) promoter in the binary expression vector, pBI1400-HN-F, and introduced into rapeseed (Brassica napus L.) using Agrobacterium-mediated transformation. The amount of the HN-F glycoprotein was estimated up to 0.18% and 0.11% of the total soluble protein (TSP) in transgenic seeds and leaves of canola, respectively. Confirmatory analyses of 36 transgenic lines revealed that the HN-F gene was integrated into the genome. Subsequently, HN-F protein could be expressed and accumulated in the seed tissue. Specific pathogen-free (SPF) chickens immunized orally with recombinant HN-F showed a significant rise in specific and hemagglutination inhibition (HI) antibodies 35 to 42 days post the first administration. The results implied the potential of transgenic canola seed-based expression for oral delivery of NDV immunogenic glycoproteins.

摘要

新城疫(ND)被认为是针对家禽的最具破坏性的传染病之一,对商业家禽生产有相当大的威胁。两种表面糖蛋白,血凝素-神经氨酸酶(HN)和融合(F),作为病毒结构中的抗原,在感染宿主细胞中也起着重要作用。在本研究中,研究了菜籽油种子中嵌合 HN-F 蛋白的表达及其在鸡中的免疫原性。HN-F 基因在二元表达载体 pBI1400-HN-F 中克隆在下脂肪酸延长酶 1(FAE1)启动子的下游,并通过根癌农杆菌介导的转化引入油菜(甘蓝型油菜)。在转基因种子和油菜叶片中,HN-F 糖蛋白的含量分别估计高达总可溶性蛋白(TSP)的 0.18%和 0.11%。对 36 个转基因系的确认分析表明,HN-F 基因已整合到基因组中。随后,HN-F 蛋白可以在种子组织中表达和积累。用重组 HN-F 经口免疫的无特定病原体(SPF)鸡在第一次给药后 35 至 42 天内特异性和血凝抑制(HI)抗体显著升高。结果表明,基于转基因油菜种子的表达系统具有口服传递 NDV 免疫原性糖蛋白的潜力。

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