Construction of lentiviral vector containing mouse aquaporin-1 gene and its expression in Schwann cells
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Supported by National Natural Science Foundation of China (31271264) and Key Basic Research Program of Shanghai Science and Technology Commission (08JC1407100).

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    Abstract:

    Objective To construct a lentiviral vector carrying mouse aquaporin-1 (AQP1) gene and use it for infecting Schwann cells of C57BL/6 mouse, so as to provide AQP1+Schwann cells for further studying the relationship of AQP1 with peripheral nerve system injury edema.Methods Mouse AQP1 gene was inserted into lentiviral vector pCDH-CMV-MCS-EF1-copGFP, and the products were confirmed by PCR and sequencing analysis. pCDH-CMV-MCS-EF1-copGFP-AQP1 and the virus packaging plasmids psPAX2 and pMD were cotransducted into 293T cells to prepare lentivirus-AQP1, and the latter was used to infect C57BL/6 mouse Schwann cells in vitro. The expression of AQP1 mRNA and protein was detected by quantitative real-time PCR and Western blotting analysis in infected mouse Schwann cells. Results PCR and sequencing revealed that the pCDH-CMV-MCS-EF1-copGFP-AQP1 plasmids were successfully constructed. The expression of AQP1 mRNA and protein in Schwann cells was significantly increased than that in cells infected with control lentiviruses (P<0.05). Conclusion We have successfully constructed a recombinant lentivirus carrying AQP1 gene, which can be used to infect mouse Schwann cells, leading to increase of AQP1 mRNA and protein expression.

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History
  • Received:September 06,2012
  • Revised:October 11,2012
  • Adopted:December 07,2012
  • Online: February 22,2013
  • Published:
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