| 拟南芥转录因子NF-YC的原核表达及多克隆抗体制备 |
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| 引用本文： | 卫静,李长江,刘亚静,黄桂雪,张凌云. 拟南芥转录因子NF-YC的原核表达及多克隆抗体制备[J]. 生物技术通报, 2012, 0(3): 57-62 |
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| 作者姓名： | 卫静 李长江 刘亚静 黄桂雪 张凌云 |
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| 作者单位： | 北京林业大学森林培育与保护教育部重点实验室,北京,100083 |
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| 基金项目： | 国家转基因生物新品种培育科技重大专项 |
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| 摘 要： | 采用PCR方法扩增NF-YC基因得到其全长cDNA序列,并将其克隆至原核表达载体pET-48b中,在大肠杆菌BL21中用IPTG诱导出分子量约为45 kD的融合蛋白,SDS-PAGE和Western blotting检测鉴定表达产物。利用亲和层析技术对融合蛋白进行纯化,纯化后的目的蛋白免疫新西兰兔制备多克隆抗体。间接ELISA检测抗体效价大于1 62 500,Western blotting结果显示,该抗体可特异性识别NF-YC蛋白。
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| 关 键 词： | 拟南芥 转录因子 NF-YC融合蛋白 原核表达 多克隆抗体 |
Prokaryotic Expression of the Arabidopsis Transcription Factor NF-YC and Preparation of Its Polyclonal Antibodies |
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| Wei Jing , Li Changjiang , Liu Yajing , Huang Guixue , Zhang Lingyun. Prokaryotic Expression of the Arabidopsis Transcription Factor NF-YC and Preparation of Its Polyclonal Antibodies[J]. Biotechnology Bulletin, 2012, 0(3): 57-62 |
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| Authors: | Wei Jing Li Changjiang Liu Yajing Huang Guixue Zhang Lingyun |
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| Affiliation: | Wei Jing Li Changjiang Liu Yajing Huang Guixue Zhang Lingyun(Key Laboratory of Forest Silviculture and Conservation of the Ministry of Education,Beijing Forestry University,Beijing 100083) |
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| Abstract: | The cDNA encoding full-length of NF-YC was amplified by PCR and was inserted into prokaryotic expression vector pET-48b.The recombinant plasmid NF-YC-pET48b was then transformed into E.coli BL21 and the NF-YC recombinant protein of 45 kD was expressed by induction with IPTG.The expression product was indentified by SDS-PAGE and Western blotting analysis.The fusion protein was purified by affinity chromatography and then was used to immunize rabbits to generate polyclonal antibody.Indirect ELISA analysis shows that the titer of the antibody is higher than 162 500.Besides,Western blotting analysis indicates that the antibody can detect expressed antigen NF-YC protein. |
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