| EGFR二聚化B细胞表位抗原肽基因克隆、表达与纯化 |
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| 引用本文： | 朱磊,吴梅芝,赵林,黄绮玲,梁梅,李黄金.EGFR二聚化B细胞表位抗原肽基因克隆、表达与纯化[J].免疫学杂志,2011(10):918-920. |
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| 作者姓名： | 朱磊 吴梅芝 赵林 黄绮玲 梁梅 李黄金 |
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| 作者单位： | 广东药学院生命科学与生物制药学院生物技术系; |
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| 基金项目： | 广东省科技计划项目(2011B080701004) |
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| 摘 要： | 目的利用基因工程手段建立表皮生长因子受体(EGFR)二聚化B细胞表位抗原肽MVF-ER的高效制备方法。方法采用重叠延伸PCR扩增MVF-ER后克隆于表达载体pET32a,于大肠杆菌BL21(DE3)进行表达,产物采用肠激酶酶切和镍离子螯合亲和层析法纯化。结果通过10对引物5步重叠延伸得到273 bp的扩增产物,目的基因与硫氧环蛋白(Trx)融合后可高效表达,经酶切和层析后得到纯度为95%的抗原肽。结论成功建立抗原肽"MVF-ER"的高效制备方法,为进一步研究EGFR过表达恶性肿瘤的治疗性疫苗打下了坚实基础。
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| 关 键 词： | 表皮生长因子受体 二聚化 B细胞表位 抗原肽 高效制备 |
Cloning,expression and purification of B cell epitope antigen peptide of EGFR dimerization |
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| ZHU Lei,WU Meizhi,ZHAO Lin,HUANG Qiling,LIANG Mei,LI Huangjin.Cloning,expression and purification of B cell epitope antigen peptide of EGFR dimerization[J].Immunological Journal,2011(10):918-920. |
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| Authors: | ZHU Lei WU Meizhi ZHAO Lin HUANG Qiling LIANG Mei LI Huangjin |
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| Institution: | ZHU Lei,WU Meizhi,ZHAO Lin,HUANG Qiling,LIANG Mei,LI HuangjinDepartment of Biotechnology,School of Life Science and Biopharmaceutics,GuangDong Pharmaceutical University,Guangzhou 510006,China |
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| Abstract: | We aim to establish a high performance preparation system for B cell epitope antigen peptide of EGFR dimerization for further research on therapeutic vaccine of EGFR over-expression tumors.The MVF-ER gene of 273 bp was amplified by splicing overlap extension PCR(SOE-PCR),then cloned into the vector of pET32a,and expressed in E.coli BL21(DE3) in the form of fusion with Trx.The expression product was purified by Ni+ affinity chromatography.The antigen peptide was recovered from the fusion protein by enterokin... |
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| Keywords: | EGFR Dimerization B cell epitope Antigen peptide High performance preparation |
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