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Abstract Objective: To express glycosyl-phosphatidylinositol (GPI) modified Met- RANTES fusion protein on Chinese hamster ovary (CHO) cells and to develop a novel immunosuppressant GPI anchored form of Met-RANTES. Methods: The eukaryotic expression vector PEF/GPI-Met-RANTES were constructed and transfected into CHO cells by electroporation. The transfectants were selected with methotrexate (MTX). Expression of the recombinant protein was assessed by flow cytometric analysis, cell immunofluorescence staining and immunogold electron microscopy. Results: The chimeric molecules of GPI anchored form of Met-RANTES including the whole reading frame were constructed, and the sequence was identical to the designed sequence. GPI anchored form of Met-RANTES was stably expressed on CHO- DHFRˉ cells. Conclusion: A large amount of GPI modified Met-RANTES fusion protein was expressed on CHO cells. GPI anchored form of Met-RANTES will may be used as novel immunosuppressant for suppressing reaction in graft rejection.
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Received: 05 December 2005
Published: 25 June 2006
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