拟南芥开花抑制基因TFL1多克隆抗体的纯化与特异性检测

摘要

This study aimed at obtaining highly sensitive and specific TFL1 polyclonal antibody.The GST-TFL1 and His-TFL1 prokaryotic expression vectors were made and transformed into E.coli expression strain BL21.The GST-TFL1 and His-TFL1 recombinant proteins were purified successfully after IPTG induction and protein purification.The molecular weights of GST-TFL1 and His-TFL1 are 46 kD and 22 kD,respectively.The TFL1 antibody serum was immuned using His-TFL1 as the antigen.The serum was purified using GST-TFL1 recombinant protein coupled affinity purification system.The western blot assay showed that the purified TFL1 antibody could specifically identify TFL1 recombinant proteins from both E.coli and Arabidopsis.The highly sensitive and specific TFL1 polyclonal antibody purified in this study would provide a powerful tool to study the mechanisms of TFL1 repressing flowering.%为了获得敏感性和特异性高的TFL1多克隆抗体,本研究构建了GST-TFL1和His-TFL1原核表达载体并转入大肠杆菌表达菌株BL21中.经IPTG诱导和蛋白纯化,成功获得了分子量约为46 kD的GST-TFL1和22 kD的His-TFL1融合蛋白.利用纯化的融合蛋白His-TFL1作为抗原免疫,获得了TFL1的多克隆抗体血清.抗体血清经偶联了GST-TFL1融合蛋白的纯化基质进行免疫亲和纯化获得了纯化的TFL1多克隆抗体.免疫印迹检测抗体敏感性和特异性发现,纯化后的抗体不仅能特异地识别细菌内纯化的GST-TFL1和His-TFL1融合蛋白,也能与拟南芥中TFL1蛋白发生特异性反应.该研究结果为深入研究TFL1抑制植物开花的分子机制提供了有力的工具.