| Ad-sTNFRI-IgGFc腺病毒载体的构建及其转染人气道平滑肌细胞 |
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| 引用本文： | 苏瑾,尤长宣,蔡绍曦,马骊,王小宁,温茜,罗微,黄泳塔.Ad-sTNFRI-IgGFc腺病毒载体的构建及其转染人气道平滑肌细胞[J].生物工程学报,2008,24(5):774-779. |
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| 作者姓名： | 苏瑾 尤长宣 蔡绍曦 马骊 王小宁 温茜 罗微 黄泳塔 |
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| 作者单位： | 1. 南方医科大学南方医院呼吸科,广州,510515
2. 南方医科大学生物技术学院分子免疫研究所,广州,510515 |
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| 基金项目： | 国家高技术研究发展计划(863) (No. 2007AA02Z458)资助。 |
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| 摘 要： | TNF-a是一种具有广泛生物学活性的前炎症细胞因子, 参与哮喘整个病理生理过程。可溶性肿瘤坏死因子受体(sTNFR)可以拮抗肿瘤坏死因子活性, 已被用来治疗与TNF相关的炎性疾病。将sTNFRI-IgGFc基因插入腺病毒穿梭质粒pDC316, 与辅助质粒pBHGloxΔE1, 3Cre共转染HEK293细胞, 重组产生Ad-sTNFRI-IgGFc, PCR鉴定毒种正确后, 进行扩增、纯化和滴度测定, 转染人气道平滑肌细胞, 利用RT-PCR、免疫组化方法, 流式细胞仪, ELISA检测转染后细胞中sTNFRI-IgGFc的转录和表达。实验结果证明成功构建了Ad-sTNFRI-IgGFc腺病毒载体, 感染性滴度达3×1010 TCID50/mL, 200 moi转染气道平滑肌细胞阳性率达99.32%, 转染后气道平滑肌细胞在mRNA和蛋白水平均有sTNFRI-IgGFc表达。转染上清稀释64倍后仍对TNF有拮抗活性。为将表达sTNFRI-IgGFc腺病毒基因治疗哮喘的实验研究提供了基础。
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| 关 键 词： | 可溶性肿瘤坏死因子受体 腺病毒载体 人气道平滑肌细胞 支气管哮喘 |
| 收稿时间： | 2007/2/28 0:00:00 |
| 修稿时间： | 2008年2月28日 |
Construction of Adenoviral Vector Encoding Soluble Human sTNFRI-IgGFc cDNA and Its Expression in Human Airway Smooth Muscle Cells (HASMCs) |
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| Jin Su,Changxuan You,Shaoxi Cai,Li M,Xiaoning Wang,Qian Wen,Wei Luo and Yongta Huang.Construction of Adenoviral Vector Encoding Soluble Human sTNFRI-IgGFc cDNA and Its Expression in Human Airway Smooth Muscle Cells (HASMCs)[J].Chinese Journal of Biotechnology,2008,24(5):774-779. |
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| Authors: | Jin Su Changxuan You Shaoxi Cai Li M Xiaoning Wang Qian Wen Wei Luo and Yongta Huang |
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| Institution: | Department of Respiratory Diseases, Nanfang hospital, Southern Medical University, Guangzhou 510515, China;Department of Respiratory Diseases, Nanfang hospital, Southern Medical University, Guangzhou 510515, China;Department of Respiratory Diseases, Nanfang hospital, Southern Medical University, Guangzhou 510515, China;Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, China;Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, China;Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, China;Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, China;Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, China |
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| Abstract: | |
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| Keywords: | soluble tumor necrosis factor alpha receptor(sTNFR) adenovirus vector human airway smooth muscle cells(HASMCs) asthma |
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