Gruschke Steffi, Bussmann Bianca M, Reiche Sven, Jassoy Christian
Institute of Virology, Faculty of Medicine, Max Bürger Research Centre, University of Leipzig, Johannisallee 30, 04103 Leipzig, Germany.
Protein Expr Purif. 2008 Oct;61(2):138-41. doi: 10.1016/j.pep.2008.04.011. Epub 2008 May 9.
Protein tagging with a peptide is a commonly used technique to facilitate protein detection and to carry out protein purification. Flexibility with respect to the peptide tag is essential since no single tag suites all purposes. This report describes the usage of two short peptides from the SARS-associated coronavirus nucleocapsid (SARS-N) protein as protein tags. Plasmids for the generation of tagged proteins were generated by ligating synthetic oligonucleotides for the peptide-coding regions downstream of the protein coding sequence. The data show recognition of prokaryotically expressed HIV-1 Gag/p24 fusion protein by Western blot and efficient affinity purification using monoclonal antibodies against the tags. The SARS peptide antibody system described presents an alternative tagging opportunity in the growing field of protein science.
用肽对蛋白质进行标记是一种常用技术,可促进蛋白质检测并进行蛋白质纯化。肽标签的灵活性至关重要,因为没有单一的标签能适用于所有目的。本报告描述了来自严重急性呼吸综合征相关冠状病毒核衣壳(SARS-N)蛋白的两种短肽作为蛋白质标签的用法。通过将用于肽编码区的合成寡核苷酸连接到蛋白质编码序列下游,构建了用于生成标记蛋白的质粒。数据显示,通过蛋白质印迹法可识别原核表达的HIV-1 Gag/p24融合蛋白,并使用针对这些标签的单克隆抗体进行高效亲和纯化。所描述的SARS肽抗体系统为蛋白质科学这一不断发展的领域提供了另一种标记选择。