| 抗菌肽MagaininⅡ单链抗体基因的构建及其表达 |
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| 引用本文： | 胡泰山 陈海旭. 抗菌肽MagaininⅡ单链抗体基因的构建及其表达[J]. 细胞与分子免疫学杂志, 2000, 16(4): 317-320 |
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| 作者姓名： | 胡泰山 陈海旭 |
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| 作者单位： | 中国科学院上海生物工程研究中心,上海 |
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| 基金项目： | 国家高技术研究发展计划项目资助!No .101 01 0203 |
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| 摘 要： | 目的 将克隆的抗体轻重链可变区基因拼接成单链抗体基因并将其在大肠杆菌中表达。方法 通过ＲＴ－ＰＣＲ从两株抗ＭａｇａｉｎｉｎⅡ杂交瘤细胞株（２Ｄ１，３Ｆ８）中克隆出ＶＨ和ＶＬ基因，然后利用重组ＰＣＲ技术，将ＶＨ和ＶＬ基因通过柔性肽段（ＧＬＹ４Ｓｅｒ）３的Ｌｉｎｋｅｒ拼接成单链抗体基因（ＳｃＦｖ），将ＳｃＦｖ克隆到表达载体ｐＣＡＮＴＡＢ５Ｅ上并将其分别在大肠杆菌Ｅ．ｃｏｋｉ ＨＢ２１５１及ＴＧ１中进行
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| 关 键 词： | MagaininⅡ 重组PCR 单链抗体 分泌表达 抗菌肽 |
| 修稿时间： | 1999-11-08 |
Construction and expression of ScFv against antibacterial peptide Magainin II |
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| HU Tai shan,CHEN Hai xu,YANG Yun gui,QIAN You cun,CHEN Deng hong,QU Xian ming,YANG Sheng liShanghai Research Center of Biotechnology,Chinese Academy of Sciences,Shanghai ,China. Construction and expression of ScFv against antibacterial peptide Magainin II[J]. Chinese journal of cellular and molecular immunology, 2000, 16(4): 317-320 |
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| Authors: | HU Tai shan CHEN Hai xu YANG Yun gui QIAN You cun CHEN Deng hong QU Xian ming YANG Sheng liShanghai Research Center of Biotechnology Chinese Academy of Sciences Shanghai China |
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| Affiliation: | HU Tai shan,CHEN Hai xu,YANG Yun gui,QIAN You cun,CHEN Deng hong,QU Xian ming,YANG Sheng liShanghai Research Center of Biotechnology,Chinese Academy of Sciences,Shanghai 200233,China |
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| Abstract: | Aim To obtain the ScFvs by splicing the VH and VL genes and then express the ScFvs in E.coli. Methods VH and VL genes were cloned from two hybridoma cell lines 2D1 and 3F8 against Magainin II and then spliced together through a flexible linker(Gly4Ser)3 to create ScFv using recombinant PCR. The ScFv genes were then cloned into expression vector pCANTAB5E and expressed in E.coli HB2151 and TG1 respectively. Results SDS PAGE and western blot analyses showed the ScFvs were secretingly expressed in E.coli HB 2151. The result of indirect ELISA showed the soluble and phage displayed ScFv had antigen binding activity. Conclusion The successful cloning and expression of ScFv laid the foundation for the design of new antibacterial peptides. |
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| Keywords: | Magainin II Recombinant PCR ScFv Se-creting expression Phage display |
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