Identification and characterization of a varicella-zoster virus DNA-binding protein by using antisera directed against a predicted synthetic oligopeptide.
Identification and characterization of a varicella-zoster virus DNA-binding protein by using antisera directed against a predicted synthetic oligopeptide.
复制标题
通过使用针对预测的合成寡肽的抗血清来鉴定和表征水痘带状疱疹病毒 DNA 结合蛋白。
DOI：
10.1128/jvi.62.3.802-809.1988
复制
发表时间：
1988
影响因子：
5.4
通讯作者：
Ruyechan,WT
中科院分区：
文献类型：
--
作者：
Kinchington,PR;Inchauspe,G;Subak-Sharpe,JH;Robey,F;Hay,J;Ruyechan,WT
我们已经在水痘-带状疱疹病毒（VZV）感染的细胞中发现了预测编码VZV类似物的单纯疱疹病毒主要dna结合蛋白的基因产物。VZV基因的开放阅读框有可能编码一种预测分子量为132,000的蛋白质（132K蛋白）。为了检测该蛋白，合成了一个与假定的开放阅读框羧基端对应的12个氨基酸的寡肽，并用于制备兔抗血清。所得抗体在免疫印迹分析和免疫沉淀中与vzv感染细胞中发现的单个130K多肽特异性反应。合成肽的加入抑制了抗血清与130K多肽的特异性反应性。用抗血清作为探针对130K多肽进行的免疫荧光研究表明，该多肽主要位于感染细胞的细胞核内。对固定在纤维素基质上的单链DNA结合蛋白的分析表明，130K多肽的30 - 50%能够与单链DNA相互作用，这种相互作用在0.5 M NaCl中被克服。因此，我们制备了一种特异性的多克隆抗血清，该血清鉴定了一种与单纯疱疹病毒ICP8 (Vmw130) dna结合蛋白性质相似的VZV dna结合蛋白。
We have identified, in varicella-zoster virus (VZV)-infected cells, the product of the gene predicted to code for the VZV analog of the herpes simplex virus major DNA-binding protein. The open reading frame of the VZV gene has the potential to code for a protein with a predicted molecular weight of 132,000 (a 132K protein). To detect the protein, a 12-amino-acid oligopeptide corresponding to the carboxyl terminus of the putative open reading frame was synthesized and used to prepare antisera in rabbits. The resulting antibodies reacted specifically in Western immunoblot analysis and immunoprecipitation with a single 130K polypeptide found in VZV-infected cells. The specific reactivity of the antisera with the 130K polypeptide was inhibited by the addition of synthetic peptide. Immunofluorescence studies with the antisera as probe for the 130K polypeptide suggested that this peptide is located predominantly within the nuclei of infected cells. Analysis of proteins that bind to single-stranded DNA immobilized on cellulose matrices indicated that 30 to 50% of the 130K polypeptide is capable of interacting with single-stranded DNA and that this interaction is overcome with 0.5 M NaCl. Thus, we have prepared a specific polyclonal antiserum that identifies a VZV DNA-binding protein whose properties are similar to those of the herpes simplex virus ICP8 (Vmw130) DNA-binding protein.