| bFGF真核表达载体构建及表达 |
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| 引用本文： | 张丽君,丁焕文,王捷,郭勇,杨传红,冼江,郑文岭. bFGF真核表达载体构建及表达[J]. 华南理工大学学报(自然科学版), 2002, 30(8): 20-23 |
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| 作者姓名： | 张丽君 丁焕文 王捷 郭勇 杨传红 冼江 郑文岭 |
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| 作者单位： | 1. 华南理工大学,食品与生物工程学院,广东,广州,510640
2. 广州军区总医院,医学实验科,广东,广州,510010 |
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| 基金项目： | 广东省自然科学基金资助项目 (990 0 0 6 ) |
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| 摘 要： | 旨在构建可在哺乳细胞中表达碱性成纤维细胞生长因子（basic Fibroblast Growth Factor,简称bFGF）的真核表达载体，以便用于研究bFGF基因治疗在骨组织工程中的应用，应用基因重组技术，将已经克隆的bFGF基因从PBR322－bFGF载体上亚克隆到真核表达载体pcDNA3．1上，构建的重组质粒经脂质体介导转染3T3细胞，36h后观察瞬时表达情况，酶切，PCR和DNA测序结果均证实了插入片段的正确性，免疫组织化学检测bFGF的表达分泌情况，结果显示部分经转染的细胞呈阳性（棕色颗粒）。结果表明已成功构建了bFGF真核表达载体pcDNA3.1-bFGF，并且bFGF能够在3T3细胞表达。
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| 关 键 词： | bFGF基因 真核表达 DNA重组技术 转染 |
| 文章编号： | 1000-565X(2002)08-0020-04 |
Construction and Expression of bFGF Gene Eukaryotic Cell Expression Vector |
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| Zhang Li-jun Ding Huan-wen Wang Jie Guo yong Yang Chuan-hong Xian Jiang Zheng Wen-ling. Construction and Expression of bFGF Gene Eukaryotic Cell Expression Vector[J]. Journal of South China University of Technology(Natural Science Edition), 2002, 30(8): 20-23 |
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| Authors: | Zhang Li-jun Ding Huan-wen Wang Jie Guo yong Yang Chuan-hong Xian Jiang Zheng Wen-ling |
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| Affiliation: | Zhang Li-jun 1 Ding Huan-wen 2 Wang Jie 2 Guo yong 1 Yang Chuan-hong 2 Xian Jiang 2 Zheng Wen-ling 2 |
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| Abstract: | The present study is aimed at constructing a mammalian basic Fibroblast Growth Factor (bFGF)recombinant vector for application of bFGF gene therapy in bone tissue engineering. bFGF gene was subcloned into pcDNA3.1 vector from PBR322 vector by means of gene cloning to obtain pcDNA3.1-bFGF plasmid, 3T3 cells were transfected with pcDNA3.1-bFGF plasmid using Lipofectamine reagent. After 36 hours the transient expression of bFGF was detected. The correct construction of pcDNA3.1-bFGF was identified by means of restriction enzyme analysis, PCR amplication and nucleotide sequence determination. Immunohistochemisty showed parts of transfected cells expressed bFGF in cells(brown staining). The pcDNA3.1-bFGF eukaryotic expression vector was constructed successfully and bFGF can be expressed in 3T3 cells. |
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| Keywords: | bFGF gene eukaryotic expression DNA recombinant technology transfection |
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