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首页> 外文期刊>Journal of Clinical Microbiology >Procaryotic expression of phosphorylated tegument protein pp65 of human cytomegalovirus and application of recombinant peptides for immunoblot analyses.
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Procaryotic expression of phosphorylated tegument protein pp65 of human cytomegalovirus and application of recombinant peptides for immunoblot analyses.

机译：人巨细胞病毒磷酸化外皮蛋白pp65的原核表达和重组肽在免疫印迹分析中的应用。

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摘要

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.
机译：人巨细胞病毒（HCMV）的外皮中含有65道尔顿的磷酸化蛋白，称为pp65，据报道该蛋白带有明显的表位，可在自然感染期间刺激体液免疫反应。针对该蛋白的单克隆抗体用于筛选重组多肽的gt11 cDNA文库。来自纯化的λ克隆的两个DNA片段和来自基因组DNA的一个片段用于克隆在细菌高水平表达载体中。测试所得的融合蛋白与一组针对pp65的单克隆抗体和多特异性抗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具有26个氨基酸的病毒部分的重组蛋白之一中。用HCMV阳性人血清进行的免疫印迹分析表明，单独的pp65并不是血清学诊断的可靠抗原，但与其他HCMV蛋白结合使用可能非常有用。

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