Improvement and observation of immunoelectron microscopic method for the localization of frog Rana grylio virus (RGV) in infected fish cells
文献类型：期刊论文
| 作者 | Huang, Xiaohong; Zhang, Qiya |
| 刊名 | MICRON
 |
| 出版日期 | 2007 |
| 卷号 | 38期号:6页码:599-606 |
| 关键词 | immunoelectron microscopy iridovirus RGV fish cell amorphous structures |
| ISSN号 | 0968-4328 |
| 通讯作者 | Zhang, QY, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China |
| 中文摘要 | In this paper, to understand the roles of amorphous structures which were observed within the viromatrix of Rana grylio virus (RGV), an improved immunoelectron microscopy (IEM) method was developed to detect the localization of RGV in carp Epithelipma papulosum cyprinid (EPC) cells. Infected EPC cells were fixed with 4% paraformaldehyde-0.25% glutaraldehyde mixture, dehydrated completely, and embedded in LR White resin. This method allowed good ultrastructural preservation and specific labeling with anti-RGV antibodies. The results of IEM showed that colloidal gold mainly bound to the capsids of viral particles at the stage of viral assembly, while during the viral maturation colloidal gold bound to the envelop of virions. In addition, within the viromatrix, the amorphous structures, including dense floccules, membranous materials and tubules, also had strong colloidal gold signals, revealing that those amorphous structures were participated in RGV assembly. In contrast, no significant gold labeling signals were obtained in negative controls. The present study not only provided further evidence that amorphous structures within the viromatrix were involved in the process of RGV assembly, but also developed an improved IEM method for studying the interaction between iridovirus and host cells. (C) 2006 Elsevier Ltd. All rights reserved. |
| 英文摘要 | In this paper, to understand the roles of amorphous structures which were observed within the viromatrix of Rana grylio virus (RGV), an improved immunoelectron microscopy (IEM) method was developed to detect the localization of RGV in carp Epithelipma papulosum cyprinid (EPC) cells. Infected EPC cells were fixed with 4% paraformaldehyde-0.25% glutaraldehyde mixture, dehydrated completely, and embedded in LR White resin. This method allowed good ultrastructural preservation and specific labeling with anti-RGV antibodies. The results of IEM showed that colloidal gold mainly bound to the capsids of viral particles at the stage of viral assembly, while during the viral maturation colloidal gold bound to the envelop of virions. In addition, within the viromatrix, the amorphous structures, including dense floccules, membranous materials and tubules, also had strong colloidal gold signals, revealing that those amorphous structures were participated in RGV assembly. In contrast, no significant gold labeling signals were obtained in negative controls. The present study not only provided further evidence that amorphous structures within the viromatrix were involved in the process of RGV assembly, but also developed an improved IEM method for studying the interaction between iridovirus and host cells. (C) 2006 Elsevier Ltd. All rights reserved. |
| WOS标题词 | Science & Technology ; Technology |
| 学科主题 | Microscopy |
| 类目[WOS] | Microscopy |
| 研究领域[WOS] | Microscopy |
| 关键词[WOS] | HEMATOPOIETIC NECROSIS VIRUS ; PSEUDORABIES VIRUS ; DIFFERENT FIXATION ; EPOXY SECTIONS ; VACCINIA VIRUS ; IRIDOVIRUS ; CONTRAST ; PROTEIN ; LINE |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000246629400007 |
| 公开日期 | 2010-10-13 |
| 源URL | [http://ir.ihb.ac.cn/handle/152342/8580]  |
| 专题 | 水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文 |
| 作者单位 | 1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China 2.Grad Univ Chinese Acad Sci, Beijing 10039, Peoples R China |
| 推荐引用方式 GB/T 7714 | Huang, Xiaohong,Zhang, Qiya. Improvement and observation of immunoelectron microscopic method for the localization of frog Rana grylio virus (RGV) in infected fish cells[J]. MICRON,2007,38(6):599-606. |
| APA | Huang, Xiaohong,&Zhang, Qiya.(2007).Improvement and observation of immunoelectron microscopic method for the localization of frog Rana grylio virus (RGV) in infected fish cells.MICRON,38(6),599-606. |
| MLA | Huang, Xiaohong,et al."Improvement and observation of immunoelectron microscopic method for the localization of frog Rana grylio virus (RGV) in infected fish cells".MICRON 38.6(2007):599-606. |
入库方式： OAI收割
来源：水生生物研究所
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