| B7-1/GFP基因真核表达载体的构建及其在骨肉瘤细胞中的表达 |
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| 引用本文： | 宁旭,刘勇,杨述华,傅德皓. B7-1/GFP基因真核表达载体的构建及其在骨肉瘤细胞中的表达[J]. 中德临床肿瘤学杂志, 2006, 5(2): 141-143. DOI: 10.1007/s10330-005-0413-6 |
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| 作者姓名： | 宁旭 刘勇 杨述华 傅德皓 |
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| 作者单位： | Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan430022, China |
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| 基金项目： | This study was supported by a grant from the youth dawn program of Wuhan （Grant No. 20025001028）. |
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| 摘 要： | Objective： To construct eukaryotic expression vector containing B7-1/GFP geneand study its expression in osteosarcoma cell line LM8. Methods： By using gene cloning technique, eukaxyotic expression vector pEGFP-C1 was used to construct the murine B7-1 recombinant plasmid （pEGFP-C1/B7）. Recombinant plasmid was transfected into LM8 cells with liposome and was confirmed by restriction endonuclease digestion and DNA sequencing. The expression of the fusion protein was detected using fluorescence microscope and Western blot analysis. Results： The recombinant eukaryotic expression plasmid pEGFP-C1/B7 was successfully constructed, which was confirmed by DNA sequencing, RT-PGR and restriction enzymes analysis. The green fluorescent protein could be detected in the transfected LM8 with fluorescence microscope. The expected B7-1 and green fluorescent protein （GFP） fusion protein was detected by RT-PCR and Western blot. Conclusion： The eukaryotic expression vector containing B7-1/GFP gene was constructed successfully, and it could be expressed in LM8 after transfection.
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| 关 键 词： | B7基因 绿色荧光蛋白 基因重组 骨肉瘤 |
| 收稿时间： | 2005-11-28 |
| 修稿时间： | 2005-12-09 |
Construction of Eukaryotic Expression Vector ContainingB7-1/GFP Gene and Its Expression in Osteosarcoma Cell Line |
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| Xu NING,Yong LIU,Shuhua YANG,Dehao FU. Construction of Eukaryotic Expression Vector ContainingB7-1/GFP Gene and Its Expression in Osteosarcoma Cell Line[J]. The Chinese-German Journal of Clinical Oncology, 2006, 5(2): 141-143. DOI: 10.1007/s10330-005-0413-6 |
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| Authors: | Xu NING Yong LIU Shuhua YANG Dehao FU |
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| Affiliation: | (1) Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 430022 Wuhan, China |
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| Abstract: | Objective: To construct eukaryotic expression vector containing B7-1/GFP geneand study its expression in osteosarcoma cell line LM8. Methods: By using gene cloning technique, eukaryotic expression vector pEGFP-C1 was used to construct the murine B7-1 recombinant plasmid (pEGFP C1/B7). Recom-binant plasmid was transfected into LM8 cells with liposome and was confirmed by restriction endonuclease digestion and DNA sequencing. The expression of the fusion protein was detected using fluorescence microscope and Western blot analysis. Results: The recombinant eukaryotic expression plasmid pEGFP C1/B7 was successfully constructed, which was confirmed by DNA sequencing, RT-PCR and restriction enzymes analysis. The green fluorescent protein could be detected in the transfected LM8 with fluorescence microscope. The expected B7-1 and green fluorescent protein (GFP) fusion protein was detected by RT-PCR and Western blot. Conclusion: The eukaryotic expression vector containing B7-1/GFP gene was constructed successfully, and it could be expressed in LM8 after transfection. |
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| Keywords: | B7-1 gene green fluorescent protein gene recombination osteosarcoma |
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