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一种参与对凡纳滨对虾 WSSV 感染的新型表皮蛋白。

A novel cuticle protein involved in WSSV infection to the Pacific white shrimp Litopenaeus vannamei.

机构信息

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

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China; Center for Ocean Mega-Science, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao, 266071, China; University of Chinese Academy of Sciences, Beijing, 100049, China.

出版信息

Dev Comp Immunol. 2020 Jan;102:103491. doi: 10.1016/j.dci.2019.103491. Epub 2019 Sep 5.

Abstract

As the most productive crustacean species in aquaculture, Litopenaeus vannamei is seriously threatened by white spot syndrome virus (WSSV), which has caused huge economic damage in the past decades. Shrimp cuticle proteins are the important components in the frontier target tissues, including cuticle and the chitinous lining of the digestive tract. In present study, a novel cuticle protein gene, named LvCPAP1, was isolated and demonstrated to play an important role in WSSV infection. The deduced amino acid sequence of LvCPAP1 contained a signal peptide and a conserved chitin-binding domain type 2 (ChBD2). Tissue distribution analysis revealed that LvCPAP1 was predominantly expressed in epidermis and stomach. The transcription levels of LvCPAP1 in epidermis and stomach were significantly regulated upon WSSV challenge. DsRNA silencing of LvCPAP1 decreased the in vivo WSSV copy numbers and the death rate of shrimp after WSSV infection, indicating that LvCPAP1 might facilitate WSSV invasion. In addition, the interaction between LvCPAP1 and the major envelop protein VP24 of WSSV was revealed by yeast two-hybrid system and further confirmed by dot blot and pull-down assays. The present study implied that cuticle protein LvCPAP1 might favor the entry process of WSSV, which provided new clues for understanding the role of cuticle proteins during virus infection.

摘要

凡纳滨对虾是水产养殖中最具生产力的甲壳类动物物种,但它受到白斑综合征病毒(WSSV)的严重威胁,过去几十年中,该病毒已造成巨大的经济损失。虾壳蛋白是前沿靶组织(包括甲壳和消化道的几丁质衬里)中的重要组成部分。在本研究中,分离出了一种新型的壳蛋白基因,命名为 LvCPAP1,并证实其在 WSSV 感染中发挥重要作用。LvCPAP1 的推导氨基酸序列包含一个信号肽和一个保守的壳聚糖结合域类型 2(ChBD2)。组织分布分析表明,LvCPAP1 主要在表皮和胃中表达。LvCPAP1 在表皮和胃中的转录水平在 WSSV 攻击后受到显著调节。LvCPAP1 的 dsRNA 沉默降低了虾体内的 WSSV 拷贝数和 WSSV 感染后的死亡率,表明 LvCPAP1 可能促进了 WSSV 的入侵。此外,酵母双杂交系统揭示了 LvCPAP1 与 WSSV 的主要包膜蛋白 VP24 之间的相互作用,并通过点印迹和下拉测定进一步证实。本研究表明,壳蛋白 LvCPAP1 可能有利于 WSSV 的进入过程,这为了解壳蛋白在病毒感染过程中的作用提供了新的线索。

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