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白斑综合征病毒(WSSV)的包膜蛋白与凡纳滨对虾类围食膜蛋白(LvPT)相互作用。

Envelope Proteins of White Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT).

作者信息

Xie Shijun, Zhang Xiaojun, Zhang Jiquan, Li Fuhua, Xiang Jianhai

机构信息

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China.

University of Chinese Academy of Sciences, Beijing, 100049, China.

出版信息

PLoS One. 2015 Dec 21;10(12):e0144922. doi: 10.1371/journal.pone.0144922. eCollection 2015.

Abstract

White spot syndrome virus (WSSV) is a major pathogen in shrimp cultures. The interactions between viral proteins and their receptors on the surface of cells in a frontier target tissue are crucial for triggering an infection. In this study, a yeast two-hybrid (Y2H) library was constructed using cDNA obtained from the stomach and gut of Litopenaeus vannamei, to ascertain the role of envelope proteins in WSSV infection. For this purpose, VP37 was used as the bait in the Y2H library screening. Forty positive clones were detected after screening. The positive clones were analyzed and discriminated, and two clones belonging to the peritrophin family were subsequently confirmed as genuine positive clones. Sequence analysis revealed that both clones could be considered as the same gene, LV-peritrophin (LvPT). Co-immunoprecipitation confirmed the interaction between LvPT and VP37. Further studies in the Y2H system revealed that LvPT could also interact with other WSSV envelope proteins such as VP32, VP38A, VP39B, and VP41A. The distribution of LvPT in tissues revealed that LvPT was mainly expressed in the stomach than in other tissues. In addition, LvPT was found to be a secretory protein, and its chitin-binding ability was also confirmed.

摘要

白斑综合征病毒(WSSV)是对虾养殖中的主要病原体。病毒蛋白与其在前沿靶组织细胞表面的受体之间的相互作用对于引发感染至关重要。在本研究中,利用凡纳滨对虾胃和肠道获得的cDNA构建酵母双杂交(Y2H)文库,以确定包膜蛋白在WSSV感染中的作用。为此,将VP37用作Y2H文库筛选的诱饵。筛选后检测到40个阳性克隆。对阳性克隆进行分析和鉴别,随后确认两个属于围食膜蛋白家族的克隆为真正的阳性克隆。序列分析表明,这两个克隆可被视为同一基因,即凡纳滨对虾围食膜蛋白(LvPT)。免疫共沉淀证实了LvPT与VP37之间的相互作用。在Y2H系统中的进一步研究表明,LvPT还可与其他WSSV包膜蛋白如VP32、VP38A、VP39B和VP41A相互作用。LvPT在组织中的分布表明,LvPT在胃中的表达高于其他组织。此外,发现LvPT是一种分泌蛋白,并且其几丁质结合能力也得到了证实。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bf78/4686314/30eb5888e084/pone.0144922.g001.jpg

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