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篇目详细内容 |
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【篇名】 |
Regulation of exogenous bFGF gene mediated by recombinant adeno-associated virus in vitro |
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【刊名】 |
Frontiers of Medicine in China |
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【刊名缩写】 |
Front. Med. China |
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【ISSN】 |
1673-7342 |
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【EISSN】 |
1673-7458 |
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【DOI】 |
10.1007/s11684-009-0042-5 |
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【出版社】 |
Higher Education Press and Springer-Verlag |
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【出版年】 |
2009 |
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【卷期】 |
3
卷2期 |
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【页码】 |
158-163
页,共
6
页 |
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【作者】 |
Ke SONG MD;
Nianjing RAO MD;
Meiling CHEN MD;
Yingguang CAO MD;
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【关键词】 |
tetracycline regulatory system; adeno-associated virus; basic fibroblast growth factor; gene regulation |
【摘要】 |
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The regulatory effect of basic fibroblast growth factor (bFGF) mediated by recombinant adeno-associated virus (AAV) in vitro was investigated. Recombinant plasmid pAAV-S3-bFGF, and pSVneo were co-transfected into BHK-21 cells, then the recombinant AAV genome was replicated and packaged with the helper virus HSV1-rc/ΔUL2. The titer of the recombinant rAAV2-tet-off-bFGF was determined by dot-blot assay. MC3T3-E1 cells were infected with rAAV2-tet-off-bFGF. Regulatory effects of Doxycycline (Dox) on bFGF and osteogenic factors were assayed quantitatively by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The physical particle titer of rAAV2-tet-off-bFGF successfully constructed was 1.8×1012 vector genomes/mL, and the virus could infect MC3T3-E1 cells effectively. In MC3T3-E1 cells treated with Dox, the expression levels of exogenous bFGF and osteogenic factors declined to varying degrees. It was concluded that rAAV2-tet-off-bFGF could infect MC3T3 cells efficiently, and this recombinant system could be regulated successfully by Dox in vitro. |
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