The Journal of international medical research · 2020
Construction and characterization of an adenoviral vector encoding human bone morphogenetic protein-2.
СтатьяНазвание и аннотация: Перевод в очереди
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Peng Liangquan, Chen Kang, Zhu Weimin, Lu Wei, Xu Jian, Huang Yong, Kuai Shengzheng, Deng Zhenhan, Wang Daping
Аннотация
OBJECTIVES: Construction of adenoviral vectors can be complicated and time-consuming. The aim of this study was to construct an adenoviral vector expressing human bone morphogenetic protein 2 (BMP-2).
METHODS: An adenoviral vector expressing human BMP-2 was constructed using the Gateway™ technique based on site-specific recombination. Briefly, BMP-2 cDNA was obtained by polymerase chain reaction, inserted into the pMD19-T cloning vector, and subcloned into a shuttle vector. The DNA sequence encoding BMP-2 was then subcloned from pEC3.1-BMP2 into the pAd/BLOCK-iT™-DEST adenoviral vector genome, which was then linearized and used to transfect 293 cells for adenovirus packaging. Adenovirus particles were generated with a titer of 1 × 1010 infectious units/mL.
RESULTS: Potent expression of BMP-2 mRNA and protein was detected in adenovirus-infected 293 cells, confirming that the adenoviral vector encoding BMP-2 was successfully constructed.
CONCLUSIONS: This recombinant adenoviral vector encoding BMP-2 can be applied in future studies to explore the roles of BMP-2 in various cell types and tissues.
METHODS: An adenoviral vector expressing human BMP-2 was constructed using the Gateway™ technique based on site-specific recombination. Briefly, BMP-2 cDNA was obtained by polymerase chain reaction, inserted into the pMD19-T cloning vector, and subcloned into a shuttle vector. The DNA sequence encoding BMP-2 was then subcloned from pEC3.1-BMP2 into the pAd/BLOCK-iT™-DEST adenoviral vector genome, which was then linearized and used to transfect 293 cells for adenovirus packaging. Adenovirus particles were generated with a titer of 1 × 1010 infectious units/mL.
RESULTS: Potent expression of BMP-2 mRNA and protein was detected in adenovirus-infected 293 cells, confirming that the adenoviral vector encoding BMP-2 was successfully constructed.
CONCLUSIONS: This recombinant adenoviral vector encoding BMP-2 can be applied in future studies to explore the roles of BMP-2 in various cell types and tissues.