| 拟南芥开花抑制基因TFL1多克隆抗体的纯化与特异性检测 |
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| 引用本文： | 袁敏,张越,王莉,葛伟娜,张岚,邢继红.拟南芥开花抑制基因TFL1多克隆抗体的纯化与特异性检测[J].河北农业大学学报,2017,40(2). |
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| 作者姓名： | 袁敏 张越 王莉 葛伟娜 张岚 邢继红 |
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| 作者单位： | 1. 华北理工大学生命科学学院/基因组学和计算生物学研究中心,河北 唐山,063000;2. 南京农业大学植物保护学院,江苏 南京,210000;3. 河北农业大学 真菌毒素与植物分子病理学实验室,河北 保定,071001 |
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| 基金项目： | 国家自然科学基金，河北省自然科学基金 |
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| 摘 要： | 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46kD的GST-TFL1和22kD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。
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| 关 键 词： | TFL1 多克隆抗体 亲和纯化 特异性 敏感性 |
Purification of flowering repressive gene TFL1 Polyclonal antibody and specificity detection in Arabidopsis thaliana |
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| YUAN Min,ZHANG Yue,WANG Li,GE Wei-na,ZHANG Lan,XING Ji-hong.Purification of flowering repressive gene TFL1 Polyclonal antibody and specificity detection in Arabidopsis thaliana[J].Journal of Agricultural University of Hebei,2017,40(2). |
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| Authors: | YUAN Min ZHANG Yue WANG Li GE Wei-na ZHANG Lan XING Ji-hong |
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| Abstract: | 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. |
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| Keywords: | TFL1 polyclonal antibody affinity purification specificity sensitivity |
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