【作者】 万晴姣； 周胜； 黄友华； 黄晓红； 曹剑豪； 秦启伟；

【Author】 Qingjiao Wan,Sheng Zhou,Youhua Huang,Xiaohong Huang,Jianhao Cao,Qiwei Qin~* (state Key Laboratory for Biocontrol,College of Life Sciences,Sun Yat-sen University,Guangzhou,China)

【机构】 中山大学生命科学学院有害生物控制与资源利用国家重点实验室；

【摘要】 新加坡石斑鱼虹彩病毒(SGIV)是一种重要的鱼类传染性病毒,可导致石斑鱼死亡率达90%以上,给石斑鱼的养殖造成巨大的经济损失。全基因组序列测定及诠释揭示SGIV为虹彩病毒科蛙病毒属的一个新种。本研究从SGIV全基因组中克隆到ORF038全长,进行原核表达,并免疫小鼠制备抗体。通过RT-PCR、Western-blot分析,结合药物抑制实验,揭示ORF038是一个晚期基因。构建真核表达重组质粒pEGFP-038并转染GP细胞,结果显示主要表达在GP细胞的细胞质中;免疫荧光显微镜观察结果高度一致。Western-blot和免疫电镜结果表明VP38定位于病毒核衣壳。抗体中和实验揭示VP38在病毒感染过程中可能起重要作用。更多还原

【Abstract】 Singapore grouper iridovirus(SGIV) belongs to the genus Ranavirus in the family Iridoviridae,and its genome was completely sequenced in 2004.The virus caused more than 90%mortality in cultured grouper.In order to better understand the viral structure and functional genes involved in infection and virus-host interactions,a candidate gene,VP38 (ORF038L) was investigated in our study.VP38 were cloned and expressed in the pET32a(+) vector,and then specific antibody was prepared.RT-PCR and Western-blot detection revealed that the VP38 transcription and translation appeared late during infection of grouper embryonic cells(GP cells).VP38 expression was also blocked by viral DNA replication inhibitor, indicating that VP38 is a late expression gene.Immunofluorescence localization found an association of ORF 038L with cytoplasm in SGIV-infected GP cells,and this association was further confirmed by observing the green fluorescence in pEGFP-VP38 transfected cells. Western blot analysis and the immunoelectron microscope method(IEM) confirmed that VP38 was present exclusively in the viral capsid fraction.The neutralization experiment suggested that VP38 might play an important role in SGIV infectivity.更多还原