• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

利用单克隆抗体对1型人类免疫缺陷病毒逆转录酶进行特性分析：C末端在抗体反应性及酶功能中的作用

Characterization of human immunodeficiency virus type 1 reverse transcriptase by using monoclonal antibodies: role of the C terminus in antibody reactivity and enzyme function.

作者信息

Tisdale M, Ertl P, Larder B A, Purifoy D J, Darby G, Powell K L

机构信息

Wellcome Research Laboratories, Beckenham, Kent, United Kingdom.

出版信息

J Virol. 1988 Oct;62(10):3662-7. doi: 10.1128/JVI.62.10.3662-3667.1988.

DOI:10.1128/JVI.62.10.3662-3667.1988
PMID:2458486
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC253508/
Abstract

We describe the production of eight monoclonal antibodies reactive with human immunodeficiency virus type 1 reverse transcriptase (RT) by immunization of mice with purified recombinant RT. These antibodies were found to react with one or the other of two regions of the enzyme and were found to be useful in immunodeficiency purification of large amounts of the enzyme. One epitope located at the C terminus of the enzyme was of particular interest, since it was present in only the larger, 66-kilodalton (kDa) RT species and not its smaller, 51-kDa counterpart. To define this epitope, a series of mutants was made which synthesized C-terminally truncated RT. These mutants indicated that the same region of the enzyme, when deleted, both removed the C-terminal epitope and drastically reduced RT activity, indicating the importance of this region in the function of the enzyme; however, even the 51-kDa enzyme component had demonstrable activity.

摘要

我们描述了通过用纯化的重组人免疫缺陷病毒1型逆转录酶（RT）免疫小鼠来产生8种与该酶反应的单克隆抗体。发现这些抗体与该酶的两个区域中的一个或另一个发生反应，并发现它们可用于大量该酶的免疫纯化。位于该酶C末端的一个表位特别令人感兴趣，因为它仅存在于较大的66千道尔顿（kDa）RT物种中，而不存在于较小的51-kDa对应物中。为了确定这个表位，制备了一系列合成C末端截短RT的突变体。这些突变体表明，该酶的同一区域在缺失时，既去除了C末端表位，又大幅降低了RT活性，表明该区域在酶功能中的重要性；然而，即使是51-kDa的酶组分也具有可证明的活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/ebcd4979faea/jvirol00089-0134-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/d21928850e79/jvirol00089-0131-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/2c2cd51cdfcc/jvirol00089-0132-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/6113660dabf6/jvirol00089-0132-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/e77f9fde92db/jvirol00089-0132-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/5ff086a8941f/jvirol00089-0133-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/ebcd4979faea/jvirol00089-0134-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/d21928850e79/jvirol00089-0131-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/2c2cd51cdfcc/jvirol00089-0132-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/6113660dabf6/jvirol00089-0132-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/e77f9fde92db/jvirol00089-0132-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/5ff086a8941f/jvirol00089-0133-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da5b/253508/ebcd4979faea/jvirol00089-0134-a.jpg

相似文献

1
Characterization of human immunodeficiency virus type 1 reverse transcriptase by using monoclonal antibodies: role of the C terminus in antibody reactivity and enzyme function.利用单克隆抗体对1型人类免疫缺陷病毒逆转录酶进行特性分析：C末端在抗体反应性及酶功能中的作用
J Virol. 1988 Oct;62(10):3662-7. doi: 10.1128/JVI.62.10.3662-3667.1988.
2
Monoclonal antibodies define linear and conformational epitopes of HIV-1 pol gene products.
AIDS Res Hum Retroviruses. 1991 Mar;7(3):307-13. doi: 10.1089/aid.1991.7.307.
3
Immunological characterization of the human immunodeficiency virus type 1 reverse transcriptase protein by the use of monoclonal antibodies.利用单克隆抗体对人类免疫缺陷病毒1型逆转录酶蛋白进行免疫学特性分析。
J Gen Virol. 1991 Aug;72 ( Pt 8):1913-8. doi: 10.1099/0022-1317-72-8-1913.
4
Structure-function relationships of HIV-1 reverse transcriptase determined using monoclonal antibodies.
J Biol Chem. 1992 Jul 25;267(21):14654-61.
5
Epitope mapping of HIV-1 reverse transcriptase with monoclonal antibodies that inhibit polymerase and RNase H activities.
J Acquir Immune Defic Syndr (1988). 1992;5(7):647-57.
6
Isolation and characterization of monoclonal antibodies raised against the reverse transcriptase of human immunodeficiency virus type 2 and cross-reactivity with that of type 1.
J Acquir Immune Defic Syndr (1988). 1993 Nov;6(11):1187-93.
7
Monoclonal antibodies against RNA-dependent DNA polymerase from myeloblastosis-associated viruses.针对成髓细胞增多症相关病毒的RNA依赖性DNA聚合酶的单克隆抗体。
Folia Biol (Praha). 1987;33(2):87-92.
8
Characterization of human immunodeficiency virus gag/pol gene products expressed by recombinant vaccinia viruses.重组痘苗病毒表达的人类免疫缺陷病毒gag/pol基因产物的特性分析
Virology. 1988 Oct;166(2):339-49. doi: 10.1016/0042-6822(88)90504-1.
9
An immunodominant neutralization epitope on the 'thumb' subdomain of human immunodeficiency virus type 1 reverse transcriptase revealed by phage display antibodies.通过噬菌体展示抗体揭示的人类免疫缺陷病毒1型逆转录酶“拇指”亚结构域上的一个免疫显性中和表位。
J Gen Virol. 2001 Apr;82(Pt 4):813-820. doi: 10.1099/0022-1317-82-4-813.
10
Production of monoclonal antibodies to human immunodeficiency virus type-2.抗人类免疫缺陷病毒2型单克隆抗体的制备。
P R Health Sci J. 1992 Dec;11(3):129-34.

引用本文的文献

1
An Alternative HIV-1 Non-Nucleoside Reverse Transcriptase Inhibition Mechanism: Targeting the p51 Subunit.一种 HIV-1 非核苷类逆转录酶抑制作用的新机制：靶向 p51 亚基。
Molecules. 2020 Dec 13;25(24):5902. doi: 10.3390/molecules25245902.
2
The cellular antiviral protein APOBEC3G interacts with HIV-1 reverse transcriptase and inhibits its function during viral replication.细胞抗病毒蛋白 APOBEC3G 与 HIV-1 逆转录酶相互作用，并在病毒复制过程中抑制其功能。
J Virol. 2012 Apr;86(7):3777-86. doi: 10.1128/JVI.06594-11. Epub 2012 Feb 1.
3
The mutation T477A in HIV-1 reverse transcriptase (RT) restores normal proteolytic processing of RT in virus with Gag-Pol mutated in the p51-RNH cleavage site.

本文引用的文献

1
Herpes simplex virus non-structural proteins. IV. Purification of the virus-induced deoxyribonuclease and characterization of the enzyme using monoclonal antibodies.单纯疱疹病毒非结构蛋白。IV. 病毒诱导的脱氧核糖核酸酶的纯化及使用单克隆抗体对该酶的特性鉴定
J Gen Virol. 1983 Oct;64 (Pt 10):2249-60. doi: 10.1099/0022-1317-64-10-2249.
2
All eight unassigned reading frames of mouse mitochondrial DNA are expressed.小鼠线粒体DNA的所有八个未分配阅读框均有表达。
EMBO J. 1984 Dec 20;3(13):3165-75. doi: 10.1002/j.1460-2075.1984.tb02275.x.
3
Monoclonal antibodies to herpes simplex virus thymidine kinase.
HIV-1 逆转录酶（RT）中的突变 T477A 恢复了在 p51-RNH 切割位点发生 Gag-Pol 突变的病毒中 RT 的正常蛋白水解加工。
Retrovirology. 2010 Feb 1;7:6. doi: 10.1186/1742-4690-7-6.
4
Virion instability of human immunodeficiency virus type 1 reverse transcriptase (RT) mutated in the protease cleavage site between RT p51 and the RT RNase H domain.在人类免疫缺陷病毒1型逆转录酶（RT）中，于RT p51和RT核糖核酸酶H结构域之间的蛋白酶切割位点发生突变后的病毒粒子不稳定性。
J Virol. 2005 Sep;79(18):11952-61. doi: 10.1128/JVI.79.18.11952-11961.2005.
5
Human recombinant antibody fragments neutralizing human immunodeficiency virus type 1 reverse transcriptase provide an experimental basis for the structural classification of the DNA polymerase family.中和人免疫缺陷病毒1型逆转录酶的人重组抗体片段为DNA聚合酶家族的结构分类提供了实验依据。
J Virol. 1996 Nov;70(11):7706-12. doi: 10.1128/JVI.70.11.7706-7712.1996.
6
Characterization of HIV-1 reverse transcriptase with antibodies indicates conformational differences between the RNAse H domains of p 66 and p 15.利用抗体对HIV-1逆转录酶进行表征，结果表明p66和p15的核糖核酸酶H结构域之间存在构象差异。
Arch Virol. 1993;131(3-4):393-403. doi: 10.1007/BF01378640.
7
Mechanism of resistance of human immunodeficiency virus type 1 to 2',3'-dideoxyinosine.1型人类免疫缺陷病毒对2'，3'-双脱氧肌苷的耐药机制。
Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6135-9. doi: 10.1073/pnas.90.13.6135.
8
Linker insertion mutagenesis of the human immunodeficiency virus reverse transcriptase expressed in bacteria: definition of the minimal polymerase domain.在细菌中表达的人类免疫缺陷病毒逆转录酶的接头插入诱变：最小聚合酶结构域的定义
Proc Natl Acad Sci U S A. 1989 May;86(9):3104-8. doi: 10.1073/pnas.86.9.3104.
9
Expression and biochemical characterization of human immunodeficiency virus type 1 nef gene product.人类免疫缺陷病毒1型nef基因产物的表达及生化特性分析
J Virol. 1990 Jul;64(7):3447-54. doi: 10.1128/JVI.64.7.3447-3454.1990.
10
Subunit-selective mutagenesis indicates minimal polymerase activity in heterodimer-associated p51 HIV-1 reverse transcriptase.亚基选择性诱变表明，在异二聚体相关的p51 HIV-1逆转录酶中，聚合酶活性极低。
EMBO J. 1991 Dec;10(12):3905-11. doi: 10.1002/j.1460-2075.1991.tb04960.x.
J Gen Virol. 1984 Sep;65 ( Pt 9):1625-30. doi: 10.1099/0022-1317-65-9-1625.
4
Virus-specific proteins synthesized in encephalomyocarditis virus-infected HeLa cells.在脑心肌炎病毒感染的HeLa细胞中合成的病毒特异性蛋白。
Proc Natl Acad Sci U S A. 1971 Dec;68(12):3083-7. doi: 10.1073/pnas.68.12.3083.
5
Rapid and efficient site-specific mutagenesis without phenotypic selection.无需表型筛选的快速高效位点特异性诱变。
Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92. doi: 10.1073/pnas.82.2.488.
6
Rapid and efficient site-specific mutagenesis without phenotypic selection.无需表型选择的快速高效位点特异性诱变。
Methods Enzymol. 1987;154:367-82. doi: 10.1016/0076-6879(87)54085-x.
7
Identification and characterization of HIV-specific RNase H by monoclonal antibody.通过单克隆抗体对HIV特异性核糖核酸酶H进行鉴定与表征
EMBO J. 1988 Jan;7(1):239-43. doi: 10.1002/j.1460-2075.1988.tb02805.x.
8
AIDS virus reverse transcriptase defined by high level expression in Escherichia coli.通过在大肠杆菌中的高水平表达所定义的艾滋病病毒逆转录酶。
EMBO J. 1987 Oct;6(10):3133-7. doi: 10.1002/j.1460-2075.1987.tb02623.x.
9
Site-specific mutagenesis of AIDS virus reverse transcriptase.艾滋病病毒逆转录酶的位点特异性诱变
Nature. 1987;327(6124):716-7. doi: 10.1038/327716a0.
10
Immunogenicity of human immunodeficiency virus (HIV) reverse transcriptase: detection of high levels of antibodies to HIV reverse transcriptase in sera of homosexual men.人类免疫缺陷病毒（HIV）逆转录酶的免疫原性：在同性恋男性血清中检测到高水平的抗HIV逆转录酶抗体。
J Clin Immunol. 1987 May;7(3):218-24. doi: 10.1007/BF00915727.