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Construction and Identification of PIRES-BMP2-TGFβ3 Bicistronic Eukayotic Expression Vector

Author: MaXiaoSong
Tutor: LiuJinZuo
School: Qingdao University
Course: Sports Medicine
Keywords: Bone morphogenetic protein- 2 Transforming growth factor β3 Eukaryotic expression vector
CLC: R87
Type: Master's thesis
Year: 2011
Downloads: 14
Quote: 0
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Abstract


The aim of the present study was designed to BMP2 and TGFβ3 gene was amplified , and build a two - gene eukaryotic expression vector containing BMP2 and TGF-β3 p Workers ERS -BMP2 - TGFβ3 for the study of the two genes were co-transfected stained mesenchymal stem cells in vivo expression , and there may be a synergistic effect to lay the foundation . First , PCR method amplified from plasmid pGEMT/BMP2 by BMP2 gene is , and even into the the double gene eukaryotic expression vector pIRES, screened positive recombinant plasmid pIRES -BMP2 ; Second , using RT-PCR method TGFβ3 gene was amplified from human embryonic tissue TGFβ3 gene into plasmid pIRES -BMP2 ; digested screened positive recombinant plasmid pIRES -BMP2 - TGFβ3 and sequencing . BMP2 and TGFβ3 gene results successfully cloned , and connect it to the destination vector pIRES double digestion and plasmid sequencing confirmed that the connections are correct , and the open reading frame of the gene is correct , pIRES - BMP2 - TGFβ3 double gene eukaryotic expression vector construction success. Conclusion The use of molecular biology techniques such as PCR, DNA recombinant successfully build a two-gene eukaryotic expression vector pIRES -BMP2 - TGFβ3 stained mesenchymal stem cells co-transfected with double genes for further study to explore the in vivo expression of inter- osteogenic differentiation of mesenchymal stem cells and synergies that may exist in the foundation .

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