Construction and biological activity study of human osteoprotegerin expressing adenoviral system

Using the isolated total RNA from osteosacoma cell line MG63, the cDNA encoding human OPG was amplified by RT-PCR. A recombinant adenoviral vector carrying cDNA of OPG was constructed and OPG expression in mouse myoblast C2C12 cells was confirmed by Western blot and ELISA. The secreted expression of...

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Published inShengwu gongcheng xuebao Vol. 19; no. 1; p. 35
Main Authors Liu, Ji-Zhong, Ji, Zong-Ling, Hu, Yun-Yu, Chen, Su-Min, Zhu, Bang-Fu, Yang, Tong-Tao
Format Journal Article
LanguageChinese
Published China 01.01.2003
Subjects
Adenoviridae - genetics
Animals
Blotting, Western
Cell Line
Enzyme-Linked Immunosorbent Assay
Genetic Vectors - genetics
Humans
Male
Mice
Mice, Inbred BALB C
Osteoclasts - metabolism
Osteoprotegerin - genetics
Osteoprotegerin - metabolism
Reverse Transcriptase Polymerase Chain Reaction
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Summary:Using the isolated total RNA from osteosacoma cell line MG63, the cDNA encoding human OPG was amplified by RT-PCR. A recombinant adenoviral vector carrying cDNA of OPG was constructed and OPG expression in mouse myoblast C2C12 cells was confirmed by Western blot and ELISA. The secreted expression of OPG protein persisted more than 6 weeks in vitro, and the growth of C2C12 cells infected by recombinant adenoviral were in good state. Osteoclasts derived from mouse bone marrow cells infected with recombinant adenoviral made less number of TRAP positive cells and resorption pits formed on dentine slices.
ISSN:1000-3061