四环素调控的介导bFGF重组腺相关病毒的构建及体外实验

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (9): 851-855.

• 基础研究论著 • 下一篇

四环素调控的介导bFGF重组腺相关病毒的构建及体外实验

宋珂,陈美玲,石琦,青莹,曹颖光^*

华中科技大学同济医学院附属同济医院口腔科湖北武汉 430030

收稿日期:2015-07-27 出版日期:2015-09-28 发布日期:2016-05-04
通讯作者: 曹颖光,电话:027-83663271
作者简介:宋珂(1982~ ),女,湖北人,博士,主治医师,主要从事口腔修复种植的临床及基础研究。
基金资助:
国家自然科学基金(编号:81400490);湖北省自然科学基金(编号: 2014CFB388)

Construction of Doxycycline-inducible Adenoviral Vector-mediated Expression of Basic Fibroblast Growth Factor in Vitro

SONG Ke, CHEN Mei-ling, SHI Qi, QING Ying, CAO Ying-guang

Department of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

Received:2015-07-27 Online:2015-09-28 Published:2016-05-04

摘要/Abstract

摘要： 目的:制备携带碱性成纤维细胞生长因子(basic fibroblast growth factor ,bFGF)的病毒载体,能被四环素调控表达。方法:构建重组质粒pAAV-S3-bFGF,生产重组病毒rAAV2-tet-off-bFGF,并感染小鼠颅骨前成骨细胞MC3T3-E1,实时定量PCR检测强力霉素(doxycycline,Dox)调控下rAAV2-tet-off-bFGF对MC3T3-E1中成骨相关因子的影响。结果:生产出高滴度及纯度的重组病毒rAAV2-tet-off-bFGF,能被Dox调控,感染 MC3T3-E1后能促进胞内成骨相关因子的表达升高。结论:这种重组病毒rAAV2-tet-off-bFGF能在Dox调控下有效传递外源基因,为骨组织工程中的种子细胞提供基因传递载体。

Abstract: Objective: To establish the dox-inducible adenoviral vector and to evaluate its ability to deliver bFGF to cells in bone tissue engineering. Methods: The adenoviral vector pAAV-S3-bFGF was constructed. Adenovirus was packaged in HEK293 cells and then infected the mouse osteoblastic cells (MC3T3-E1). The efficiency of infection was evaluated by real-time PCR and Western blot. Results: The recombinant adenovirus rAAV2-tet-off-bFGF was regulated by Dox. MC3T3-E1 cells could be efficiently infected. Additionally, the expression of osteogenesis-related genes was significantly increased in vitro. Conclusion: The rAAV2-tet-off-bFGF can deliver exogenous genes efficiently into MC3T3-E1 cells, and has the potential to be used in bone tissue engineering.

Key words: Adeno-associated virus, Gene delivery, Basic fibroblast growth factor, Tetracycline regulated system

中图分类号:

R782.1

宋珂,陈美玲,石琦,青莹,曹颖光. 四环素调控的介导bFGF重组腺相关病毒的构建及体外实验[J]. 口腔医学研究, 2015, 31(9): 851-855.

SONG Ke, CHEN Mei-ling, SHI Qi, QING Ying, CAO Ying-guang. Construction of Doxycycline-inducible Adenoviral Vector-mediated Expression of Basic Fibroblast Growth Factor in Vitro[J]. Journal of Oral Science Research, 2015, 31(9): 851-855.

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四环素调控的介导bFGF重组腺相关病毒的构建及体外实验

Construction of Doxycycline-inducible Adenoviral Vector-mediated Expression of Basic Fibroblast Growth Factor in Vitro

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