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一种与疾病相关的海龟疱疹病毒免疫原性蛋白的鉴定与表达

Identification and expression of immunogenic proteins of a disease-associated marine turtle herpesvirus.

作者信息

Coberley Sadie S, Condit Richard C, Herbst Lawrence H, Klein Paul A

机构信息

Interdisciplinary Program in Biomedical Science, College of Medicine, University of Florida, Gainesville, FL 32610, USA.

出版信息

J Virol. 2002 Oct;76(20):10553-8. doi: 10.1128/jvi.76.20.10553-10558.2002.

Abstract

Herpesviruses are associated with several diseases of marine turtles, including lung-eye-trachea disease (LETD) and fibropapillomatosis. Two approaches were used to identify immunodominant antigens of LETV, the LETD-associated herpesvirus. The first approach targeted glycoprotein B, which is known to be immunogenic and neutralizing in other species. The second strategy identified LETV proteins recognized on Western blots by antibodies in immune green turtle plasma. A 38-kDa protein was resolved by two-dimensional gel electrophoresis, sequenced, and identified as a scaffolding protein encoded by the overlapping open reading frames of UL26 and UL26.5. Glycoprotein B and the scaffolding protein were cloned and expressed in Escherichia coli. The expressed proteins were recognized on Western blots by antibodies in immune green turtle plasma. Phylogenetic studies based on UL26, DNA polymerase, and glycoprotein B revealed that LETV clusters with the alphaherpesviruses.

摘要

疱疹病毒与海龟的多种疾病有关,包括肺-眼-气管疾病(LETD)和纤维乳头瘤病。采用了两种方法来鉴定与LETD相关的疱疹病毒——LETV的免疫显性抗原。第一种方法针对糖蛋白B,已知其在其他物种中具有免疫原性和中和作用。第二种策略鉴定了免疫绿海龟血浆中的抗体在蛋白质印迹法上识别的LETV蛋白。通过二维凝胶电泳分离出一种38 kDa的蛋白质,对其进行测序,并鉴定为一种由UL26和UL26.5的重叠开放阅读框编码的支架蛋白。糖蛋白B和支架蛋白在大肠杆菌中进行克隆和表达。免疫绿海龟血浆中的抗体在蛋白质印迹法上识别出表达的蛋白质。基于UL26、DNA聚合酶和糖蛋白B的系统发育研究表明,LETV与α疱疹病毒聚类。

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