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    LI Wei, WANG Wen-ping, CHEN Liang, WANG Zhen-xiang. The construction of osteopontin gene recombinant plasmid and its package of lentivirus[J]. Journal of Bengbu Medical University, 2015, 40(5): 564-568. DOI: 10.13898/j.cnki.issn.1000-2200.2015.05.002
    Citation: LI Wei, WANG Wen-ping, CHEN Liang, WANG Zhen-xiang. The construction of osteopontin gene recombinant plasmid and its package of lentivirus[J]. Journal of Bengbu Medical University, 2015, 40(5): 564-568. DOI: 10.13898/j.cnki.issn.1000-2200.2015.05.002
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    The construction of osteopontin gene recombinant plasmid and its package of lentivirus

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    • Objective: To construct the recombinant plasmid containing the osteopontin(OPN) gene and package lentivirus with it.Methods: The total RNA of kidney tissue was abstracted from C57BL/6 wild type mouse,and then was transcribed to cDNA.The OPN gene was largely amplified with special primers,and then cloned to lentivirus carrier to constract the pCDH-CMV-MCS-EF1-GFP-T2A-Puro.The lentivirus was packaged by transfecting the plasmids into 293T cells,and the virus titer was tested.Results: The OPN gene fragment was successfully cloned by sequencing.The virus titer was 2.5×108 TU/ml.Conclusions: The lentivirus containing OPN gene is successfully packaged,which lays the foundation in further researching the biological function of OPN gene.
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