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[Construction of eukaryotic expression vector of rat HAS-3 gene and its effects on chemotaxis]
1Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China. nieyong@gmail.com
Aim:
To construct the eukaryotic expression vector of rat HAS-3 gene, express it in RSC96 cells and detect the enzyme activity of the recombinant protein.
Methods:
Rat HAS-3 gene was cloned from CCI rat injury nerve cDNA using RT-PCR and inserted into pcDNA3.1D and pEGFP-N1. The recombinant plasmids were transfected to RSC96 cells. The expression level and enzyme activities of the recombinant proteins were monitored.
Results:
Rat HAS-3 gene was cloned into pcDNA3.1D and pEGFP-N1 correctly. Recombinant proteins were detected in RSC96 cells and the synthesis of HA was up-regulated after transcfection. The condition medium of RSC96 cells overexpressing HAS-3 had some effects on chemotaxis of macrophages.
Conclusion:
The eukaryotic expression vectors of rat HAS-3 has been constructed successfully. The transfection of HAS-3 gene to RSC96 cells is effective in chemotaxis of macrophages which demonstrates HAS-3 may contribute to the development of inflammation in vivo.
