Peritrophin-like protein from Litopenaeus vannamei (LvPT) involved in white spot syndrome virus (WSSV) infection in digestive tract challenged with reverse gavage  

作　　者：谢世筠 李富花 张晓军 张继泉 相建海 

机构地区：[1]Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China [2]University of Chinese Academy of Sciences, Beijing 100049, China [3]Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China

出　　处：《Chinese Journal of Oceanology and Limnology》2017年第6期1524-1530,共7页中国海洋湖沼学报（英文版）

基　　金：Supported by the National Basic Research Program of China(973 Program)(No.2012CB114403);the China Agriculture Research System-47(CARS-47)

摘　　要：The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus （WSSV） in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of Litopenaeus vannamei （LvPT） was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of LvPT was predicted with the GenScript OptimumAntigenTM design tool. An anti-LvPT polyclonal antibody was produced and shown to specifically bind a band at -27 kDa, identified as LvPT. The LvPT protein was expressed and its concentration determined. LvPT dsRNA （4 pg per shrimp） was used to inhibit LvPT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection （hpi）. Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after LvPT dsRNA interference than in those of the controls at 48 and 72 hpi. Our results imply that LvPT plays an important role during WSSV infection of the digestive tract.The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus(WSSV) in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of L itopenaeus vannamei(Lv PT) was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of Lv PT was predicted with the Gen Script Optimum Antigen? design tool. An anti-Lv PT polyclonal antibody was produced and shown to specifically bind a band at ~27 k Da, identified as Lv PT. The Lv PT protein was expressed and its concentration determined. L v PT ds RNA(4 μg per shrimp) was used to inhibit Lv PT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection(hpi). Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after L v PT ds RNA interference than in those of the controls at 48 and 72 hpi. Our results imply that Lv PT plays an important role during WSSV infection of the digestive tract.

关 键 词：Litopenaeus vannamei digestive tract peritrophin-like protein dsRNA interference binding specificity white spot syndrome virus （WSSV） reverse gavage 

分 类 号：S945.49[农业科学—水产养殖]