Construction of adeno-associated virus system for human bone morphogenetic protein 7 gene
Construction of adeno-associated virus system for human bone morphogenetic protein 7 gene
复制标题
人骨形态发生蛋白7基因腺相关病毒系统的构建
DOI：
10.1007/s11596-008-0105-x
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发表时间：
2008-02
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
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作者：
为了构建人骨形态发生蛋白7 （human bone morphogenetic protein 7, BMP7）重组腺相关病毒（adeno-associated virus, rAAV）载体，并在体外观察其mRNA表达，将BMP7 CDS序列克隆到AAV Helper Free System的表达质粒pAAV-MCS中。重组质粒经酶切和测序鉴定。重组质粒pAAV-RC、pHelper按照磷酸钙法共转染AAV-293细胞。通过4轮冷冻/解冻收集病毒库。重组病毒纯化浓缩后，采用点印迹法测定病毒滴度。将重组病毒转染不同MOI的HEK293细胞，RT-PCR检测BMP7 mRNA的表达。结果表明，rAAV-BMP7已成功构建和封装。物理颗粒滴度为2.5×1011vector基因组/mL。转染后BMP7 mRNA表达水平不同。这些数据表明，重组AAV介导hBMP7 mRNA在293细胞中的稳定表达。该方法为修复种植体周围颌骨缺损提供了一种有效的方法。
To construct the recombinant adeno-associated virus (rAAV) vector with human bone morphogenetic protein 7 (BMP7) and observe the BMP7 mRNA expressionin vitro, BMP7 CDS sequence was cloned into expression plasmid pAAV-MCS of AAV Helper Free System. The recombinant plasmid was identified with enzyme digestion and sequencing. The recombinant plasmid, pAAV-RC, pHelper were co-transfected into AAV-293 cells according to the calcium phosphate-based protocol. The viral stock was collected by 4 rounds of freeze/thaw. After purified and concentrated, the recombinant virus titer was determined by dot-blot assay. HEK293 cells were transfected with the recombinant virus at different MOI, and the expression of BMP7 mRNA was detected by RT-PCR. The results showed rAAV-BMP7 was constructed and packaged successfully. The physical particle titer was 2.5×1011vector genomes/mL. There was different expression level of BMP7 mRNA after transfecton. These data suggested that recombinant AAV mediated a stable expression of hBMP7 mRNA in 293 cells. The AAV production method may pave the way of an effective strategy for the jaw bone defection around dental implants.
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影响因子：
5.4
作者：
T. Athanasopoulos;S. Fabb;G. Dickson
通讯作者：
T. Athanasopoulos;S. Fabb;G. Dickson
影响因子：
4.2
作者：
Stephen D. Cook;G. C. Baffes;M. M. Wolfe-M.;T. K. Sampath;D. Rueger
通讯作者：
Stephen D. Cook;G. C. Baffes;M. M. Wolfe-M.;T. K. Sampath;D. Rueger
影响因子：
5.5
作者：
G. Gallez-Hawkins;Xiuli Li;Anne E. Franck;Lia Thao;S. Lacey;D. Diamond;J. Zaia
通讯作者：
G. Gallez-Hawkins;Xiuli Li;Anne E. Franck;Lia Thao;S. Lacey;D. Diamond;J. Zaia
DOI：
--
发表时间：
2007
期刊：
Journal of Brain and Nervous Diseases
影响因子：
--
作者：
Chen Jia-feng
通讯作者：
Chen Jia-feng
影响因子：
3.7
作者：
M. Ulrich‐Vinther
通讯作者：
M. Ulrich‐Vinther