| 尘螨变应原Der f1 真核表达载体的构建及转染CHO 细胞 |
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| 引用本文： | 彭江龙,崔玉宝,钱士匀,裴华,陈年根,黄幼生.尘螨变应原Der f1 真核表达载体的构建及转染CHO 细胞[J].现代生物医学进展,2011,11(14):2612-2614. |
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| 作者姓名： | 彭江龙 崔玉宝 钱士匀 裴华 陈年根 黄幼生 |
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| 作者单位： | 1. 海南医学院,海南海口,571101
2. 盐城卫生职业技术学院,江苏盐城,224006 |
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| 基金项目： | 国家自然科学基金(30860261) |
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| 摘 要： | 目的:构建尘螨变应原Der f1真核表达载体,转染真核细胞并进行蛋白表达.方法:根据Genebank中Der f1基因的核酸序列(AB034946),设计引物,采用PCR法,从保存的JM109工程菌中扩增Der f1编码基因,克隆到真核表达质粒pcDNA3.1/myc-hisA上,以脂质体法转染CHO细胞,经G418筛选,进行稳定表达细胞株的筛选和鉴定.结果:将目的基因Der f1成功连接到pcDNA3.1/myc-hisA-Derf1并转染CHO细胞,获得稳定表达的CHO细胞株.结论:成功构建了尘螨变应原Der f1真核表达载体,并转染CHO细胞表达蛋白质.
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| 关 键 词： | 尘螨 Der f1基因 真核转染 CHO细胞 |
Construction of Eukaryotic Vector of Derf1 Gene of Dermatophagoides
Farinae and Transfection of CHO Cells |
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| PENG Jiang-long,CUI Yu-bao,QIAN Shi-yun,PEI Hu,CHEN Nian-gen,HUANG You-sheng,HUANG Mian-qing.Construction of Eukaryotic Vector of Derf1 Gene of Dermatophagoides
Farinae and Transfection of CHO Cells[J].Progress in Modern Biomedicine,2011,11(14):2612-2614. |
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| Authors: | PENG Jiang-long CUI Yu-bao QIAN Shi-yun PEI Hu CHEN Nian-gen HUANG You-sheng HUANG Mian-qing |
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| Institution: | PENG Jiang-long1,CUI Yu-bao2,QIAN Shi-yun1,PEI Hua1,CHEN Nian-gen1,HUANG You-sheng1,HUANG Mian-qing1(1 Hainan Medical College,Haikou 571101,China,2 Yancheng Health Vocational & Technical College,Yancheng 224006,China) |
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| Abstract: | Objective: To construct eukaryotic expressing vector of derf1 gene of dermatophagoides farinae,and transfect CHO cells for protein expression.Methods: According to the Genebank nucleic acid sequences of Derf1(No.AB034946),designed primers,amplified the Der f 1 gene from the preservative engineering bacteria JM109 by PCR,then cloned it into plasmid pcDNA3.1/myc-his A;and then transfected the plasmid into CHO cells by liposomes and screening the positive cell clone use G418 to express the protein of Derf1 gen... |
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| Keywords: | Dermatophagoides Farinae Der f1 gene Eukaryotic transfection CHO cell |
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