【摘要】 目的 :制备p6 2DokPTB结构域的GST融合蛋白。方法 :用PCR方法扩增编码p6 2DokPTB结构域的cD NA ,并将其克隆入表达载体pGEX 4T 3中 ,转化大肠杆菌BL 2 1,构建成表达GST PTB融合蛋白的菌株。经IPTG诱导表达后 ,经Glutathion Sepharose 4B亲和层析柱纯化 ,用SDS PAGE进行分析 ,该融合蛋白的表达量超过菌体总蛋白的 2 5 %。结果 :获得重组GST PTB融合蛋白 ,纯度在 95 %以上 ,产物得率约 75 %。结论 :成功制备高纯度p6 2DokPTB结构域的GST融合蛋白 ,为进一步研究p6 2DokPTB结构域的结构和生物学功能奠定了基础。更多还原

【Abstract】 Aim:To produce the GST PTB domain fusion protein Methods:The cDNA encoding p62Dok PTB domain was amplified by using PCR method, and cloned into pGEX 4T 3 vector The expression plasmid was introduced into E coli strain BL 21 The GST PTB fusion protein accounted for over 25% of the total bacterial protein after IPTG induction, It was purified to homogeneity from the cell lysates via affinity chromatography and analyzed by SDS PAGE gel Results:The recombinant GST PTB fusion proteins were prepared with high purity, more than 95%, and high recovery, about 75% Conclusion: The established method for preparation large quantity recombinant GST PTB fusion proteins would promote the structure function analysis of the PTB domain of p62 Dok protein更多还原