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人巨细胞病毒磷酸化被膜蛋白pp65的原核表达及重组肽在免疫印迹分析中的应用

Procaryotic expression of phosphorylated tegument protein pp65 of human cytomegalovirus and application of recombinant peptides for immunoblot analyses.

作者信息

Plachter B, Klages S, Hagelmann S, Britt W, Landini M P, Jahn G

机构信息

Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, Federal Republic of Germany.

出版信息

J Clin Microbiol. 1990 Jun;28(6):1229-35. doi: 10.1128/jcm.28.6.1229-1235.1990.

DOI:10.1128/jcm.28.6.1229-1235.1990
PMID:2166078
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC267910/
Abstract

The tegument of human cytomegalovirus (HCMV) contains a phosphorylated protein of 65 kilodaltons, termed pp65, which was reported to carry significant epitopes for the stimulation of the humoral immune response during natural infection. A monoclonal antibody directed against this protein was used to screen a lambda gt11 cDNA library for recombinant polypeptides. Two DNA fragments from purified lambda clones and one fragment from genomic DNA were used for cloning in a bacterial high-level expression vector. The resulting fusion proteins were tested for their reactivity with a panel of monoclonal antibodies directed against pp65 and with polyspecific anti-HCMV rabbit antisera. The binding site for all the monoclonal antibodies tested was found to be contained in one of the recombinant proteins with a viral portion of 26 amino acids. Immunoblot analyses with HCMV-positive human sera revealed that pp65 alone is not a reliable antigen for serodiagnosis but may be very useful in combination with other HCMV proteins.

摘要

人类巨细胞病毒（HCMV）的包膜含有一种65千道尔顿的磷酸化蛋白，称为pp65，据报道，该蛋白在自然感染期间携带刺激体液免疫反应的重要表位。一种针对该蛋白的单克隆抗体被用于筛选λgt11 cDNA文库以寻找重组多肽。来自纯化的λ克隆的两个DNA片段和来自基因组DNA的一个片段被用于在细菌高效表达载体中进行克隆。对所得融合蛋白进行检测，以观察它们与一组针对pp65的单克隆抗体以及多特异性抗HCMV兔抗血清的反应性。发现所有测试的单克隆抗体的结合位点都包含在一种重组蛋白中，该重组蛋白含有一段26个氨基酸的病毒部分。用HCMV阳性人血清进行的免疫印迹分析表明，单独的pp65不是血清学诊断的可靠抗原，但与其他HCMV蛋白联合使用可能非常有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/a2fa6f144cb4/jcm00054-0169-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/b14019e0f4a9/jcm00054-0165-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/5e7b13bb5a70/jcm00054-0167-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/0f9acca01e42/jcm00054-0167-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/1f427a737f97/jcm00054-0168-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/a2fa6f144cb4/jcm00054-0169-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/b14019e0f4a9/jcm00054-0165-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/5e7b13bb5a70/jcm00054-0167-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/0f9acca01e42/jcm00054-0167-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/1f427a737f97/jcm00054-0168-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de43/267910/a2fa6f144cb4/jcm00054-0169-a.jpg

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本文引用的文献

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