中国科学院水生生物研究所机构知识库
Advanced  
IHB OpenIR  > 中科院水生所知识产出(2009年前)  > 期刊论文
题名: Preparation of polyclonal antibody and virus-induced tissue expression of Paralichthys olivaceu HRI
作者: Zhu Rong ; Zhang Yi-Bing ; Zhang Qi-Ya ; Gui Jian-Fang
通讯作者: Zhu Rong ; Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol and Biotechnol, Wuhan 430072, Peoples R China
刊名: Acta Hydrobiologica Sinica
发表日期: 2007
卷: 31, 期:1, 页:119-124
收录类别: 其他
英文摘要: The heme-regulated initiation factor 2 alpha (eIF2 alpha) kinase (HRI) is one of Ser/Thr kinases that phosphorylate the alpha-subunit of eIF2 to inhibit protein synthesis in eukaryotic cells under various stress conditions, including heme deficiency and heat shock. HRI is firstly discovered in rabbit reticulocytes as an inhibitor of globin synthesis under heme deficiency, but so far little is known about the importance of HRI in stress conditions other than heme deficiency and the roles of HRI in nonerythroid tissues. For example, there was an argument on the tissue specificity of HRI expression due to the fact that no HRI protein expression was reported in nonerythroid tissues and a finding that in HRI knockout mice, only red blood cells (RBCs) and their precursors were directly affected by the lack of HRI. Paralichthys olivaceu HRI is the first identified fish homologue, and a previous study showed for the first time that PoHRI could be induced by virus infection in vitro. In order to further investigate whether PoHRI protein is also synthesized in nonerythroid tissues and can be induced by virus infection in vivo, here a anti-PoHRI polyclonal antibody was prepared and subsequently used to detect the expression of PoHRI in different tissues of flounder. Firstly, the cDNA fragment encoding 1-200 amino acids of PoHRI was cloned and integrated into a prokaryotic expression vector pET32a. Next, a recombinant protein similar to the expected size was induced and then purified by Ni2+ -NTA chromatography. The anti-PoHRI polyclonal antibody was then prepared by immunization of mice. The specific recognition of anti-PoHRI polyclonal antibody to PoHRI was further verified by a test in which anti-PoHRI antiserum, after pre-adsorbed with recombinant PoHRI peptide, cannot recognize the corresponding polypeptide. In healthy flounder tissues, the constitutive expression of PoHRI was able to be detected at both mRNA and proteins levels, and SMRV infection was able to induce the upregulation of PoHRI in the corresponding tissues, especially in head kidney and spleen. These results support the notion that PoHRI is ubiquitously expressed in flounder tissues, which indicates that it may play an important role in cellular antiviral immune response. It is well known that PKR is the primary eIF2 alpha kinase responsive to virus infection; however, previous reports also showed that the phosphorylation of eIF2a in cultured mammalian cells in response to heat shock was apparently due to the activation of eIF2 alpha kinases including HRI and PKR, indicating that HRI and PKR were likely redundant and could functionally replace one another under some conditions. In fact, a recent study revealed that GCN2, another eIF2 alpha kinase, participate in inhibition of sindbis virus (SV) replication by phosphorylating eIF2a to block early viral translation of genomic SV RNA. However, GCN2 was originally characterized as being required for amino-acid control of GCN4 mRNA. Therefore, the upregulation of PoHRI by virus infection indicates that PoHRI may exert inhibition of protein synthesis in virus-infected flounder, a function similar to mammalian PKR. In addition, PoHRI might exhibit a new uncharacterized, heme-independent function other than translational shutoff under virus infection, since mammalian NF-kappa B can be activated by HRI through phosphorylation of inhibitor I kappa B. Therefore, further studies will be needed to understand the importance of PoHRI in flounder under virus infection.
ISSN号: 1000-3207
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/18295
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

Files in This Item: Download All
File Name/ File Size Content Type Version Access License
Preparation of polyclonal antibody and virus-induced tissue expression of Paralichthys olivaceu HRI.pdf(583KB)----开放获取--View Download

Recommended Citation:
Zhu Rong; Zhang Yi-Bing; Zhang Qi-Ya; Gui Jian-Fang.Preparation of polyclonal antibody and virus-induced tissue expression of Paralichthys olivaceu HRI,Acta Hydrobiologica Sinica,2007,31(1):119-124
Service
Recommend this item
Sava as my favorate item
Show this item's statistics
Export Endnote File
Google Scholar
Similar articles in Google Scholar
[Zhu Rong]'s Articles
[Zhang Yi-Bing]'s Articles
[Zhang Qi-Ya]'s Articles
CSDL cross search
Similar articles in CSDL Cross Search
[Zhu Rong]‘s Articles
[Zhang Yi-Bing]‘s Articles
[Zhang Qi-Ya]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
Add to CiteULike Add to Connotea Add to Del.icio.us Add to Digg Add to Reddit
文件名: Preparation of polyclonal antibody and virus-induced tissue expression of Paralichthys olivaceu HRI.pdf
格式: Adobe PDF
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Copyright © 2007-2016  中国科学院水生生物研究所 - Feedback
Powered by CSpace