Construction of lentiviral vector containing mouse aqua porin-1 gene and its expression in Schwann cells
10.3724/SP.J.1008.2013.00199
- Author:
Jie ZHANG
1
Author Information
1. Department of Plastic Surgery, Changzheng Hospital, Second Military Medical University
- Publication Type:Journal Article
- Keywords:
Aquaporin 1;
Lentivirus;
Over-expression;
Schwann cells
- From:
Academic Journal of Second Military Medical University
2013;34(2):199-204
- CountryChina
- Language:Chinese
-
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-EFl-copGFP, and the products were confirmed by PCR and sequencing analysis. pCDH-CMV-MCS-EFl-copGFP-AQPl and the virus packaging plasmids psPAX2 and pMD were cotransducted into 293T cells to prepare lentivirus-AQPl, and the latter was used to infect C57BL/6 mouse Schwann cells in vitro. The expression of AQP1 mRN A 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-EFl-copGFP-AQPl plasmids were successfully constructed. The expression of AQP1 mRNA and protein in Schwann cells was significantly increased than that in cells infected with control Antiviruses (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.
