渔业科学进展2018，Vol.39Issue(2)：138-145,8.DOI:10.19663/j.issn2095-9869.20170222002
凡纳滨对虾网格重链蛋白与WSSV结构蛋白在体外的相互作用
In vitro Interaction Between Domain of Clathrin Heavy Chain in Litopenaeus vannamei and WSSV Structural Proteins
摘要
Abstract
Clathrin heavy chain protein (CHC),a conserved protein only found in eukaryotes,is one of the essential components of clathrin.White spot syndrome virus (WSSV) infection is one of the main diseases in aquaculture.It belongs to the Whispovirus genus of the Nimaviridae family.WSSV,a rod-shaped,non-occluded baculovirus with large doubled-stranded DNA,can infect shrimps such as penaeid shrimp and crayfish.WSSV has a wide range of hosts in crustacean with high infection and mortality rate,and it causes up to 100％ mortality within 3～7 days,which causes great economic loss in aquaculture industry.Recent research showed that the clathrin mediated endocytosis path regulates WSSV infection in the Cherax quadricarinatus hematopoietic cells.However,whether the clathrin heavy chain protein plays an important way in the clathrin mediated endocytosis pathway is unclear.To address this,two primers were designed to clone the LvCHC1 and LvCHC2 according to the two structural domains:clathrin propel repeat (LvCHC1) and clathrin heavy chain repeat homology (LvCHC2) of Litopenaeus vannamei CHC.LvCHC 1 and LvCHC2 were cloned into prokaryotic expression vector pBAD/gⅢA and transformed into E.coli TOP 10.The recombinant LvCHC1 and LvCHC2 were successfully obtained by inducing with L-arabinose.The pure LvCHC1 and LvCHC2 were acquired using Co2+ affinity chromatography purification.Mass spectrometry analysis showed the correctness of the recombinant LvCHC1 and LvCHC2.Far-Western blot results indicated that LvCHC1 and LvCHC2 interacted with VP26 and VP37 with higher banding activity with VP26,and that LvCHC1 and LvCHC2 did not bind to VP28N.Taken together,the results indicated that clathrin heavy chain-mediated endocytosis is required for WSSV infection.关键词
凡纳滨对虾/网格重链蛋白/WSSV/体外作用Key words
Litopenaeus vannamei/Clathrin heavy chain protein/White Spot Syndrome Virus/Interaction in vitro分类
农业科技引用本文复制引用
王中一,刘庆慧,黄倢..凡纳滨对虾网格重链蛋白与WSSV结构蛋白在体外的相互作用[J].渔业科学进展,2018,39(2):138-145,8.基金项目
国家自然科学基金(31672679)、国家重点基础研究发展计划(2012CB114401)和泰山学者“建设工程专项经费”共同资助 [This work was supported by National Science Foundation of China (31672679),National Key Basic Research Development Plan (2012CB114401),and Taishan Scholar "Special Funds to Construction Projects"]. （31672679）
