| 彭江龙1 崔玉宝2 钱士匀1 裴华1 陈年根1 黄幼生1.尘螨变应原Der f1 真核表达载体的构建及转染CHO 细胞[J].,2011,11(14):2612-2614 |
| 尘螨变应原Der f1 真核表达载体的构建及转染CHO 细胞 |
| Construction of Eukaryotic Vector of Derf1 Gene of DermatophagoidesFarinae and Transfection of CHO Cells |
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| DOI： |
| 中文关键词: 尘螨 Der f1 基因 真核转染 CHO 细胞 |
| 英文关键词: Dermatophagoides Farinae Der f1 gene Eukaryotic transfection CHO cell |
| 基金项目:国家自然科学基金(30860261) |
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| 中文摘要: |
| 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-his A 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。 |
| 英文摘要: |
| 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 gene. Results: The Derf1 gene was connected to plasmid pcDNA3.1/myc-hisA and screening into CHO eukaryotic expression cells
successfully. Conclusions: The eukaryotic expression vector of Derf1 was constructed well and got its protein. |
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