精品文档---下载后可任意编辑DEK 蛋白的表达和纯化及其功能的初步讨论的开题报告摘要：DEK 蛋白是一种高度保守的核内 DNA 结合蛋白，参加多种细胞过程如转录和DNA 损伤修复。近年来，DEK 蛋白在肿瘤发生、进展和预后方面的讨论越来越受到重视。本讨论旨在构建 DEK 蛋白表达和纯化系统，并初步探究其功能。首先，利用 PCR扩增人 DEK 基因并克隆到表达载体 pET28a(+)，然后通过大肠杆菌 BL21 (DE3)细胞表达及纯化获得重组 DEK 蛋白。接着，利用 GST pull down 和电泳移位实验初步探究 DEK 蛋白在 DNA 结合及与其他蛋白相互作用中的作用。最后，利用小鼠淋巴瘤细胞株 EL4 建立 DEK 蛋白稳定表达细胞系，从而进一步讨论其在 DNA 损伤和肿瘤生物学中的功能。关键词：DEK 蛋白；表达；纯化；功能Abstract:DEK protein is a highly conserved nuclear DNA binding protein that is involved in various cellular processes such as transcription and DNA damage repair. In recent years, research on the role of DEK protein in tumor occurrence, progression, and prognosis has been increasingly valued. The aim of this study is to construct a DEK protein expression and purification system and to explore its function preliminarily. Firstly, the human DEK gene was amplified by PCR and cloned into the expression vector pET28a(+), and the recombinant DEK protein was obtained by expression and purification in Escherichia coli BL21 (DE3) cells. Then, the role of DEK protein in DNA binding and interaction with other proteins was preliminarily explored using GST pull-down and electrophoretic mobility shift assay. Finally, a stable cell line expressing DEK protein was established in mouse lymphoma cell line EL4 to further investigate its function in DNA damage and tumor biology.Keywords: DEK protein; expression; purification; function
