Construction and Eukaryotic Expressing of GPI-Anchored Met-RANTES
- VernacularTitle:GPI锚固型Met-RANTES真核表达载体的构建和表达
- Author:
Zhen LI
;
Feng XIA
;
Yuan-Qiang ZHANG
;
- Publication Type:Journal Article
- Keywords:
GQI-anchor Met-RANTES CHO-DHFR- cell Eukaryotic expression
- From:
China Biotechnology
2006;0(06):-
- CountryChina
- Language:Chinese
-
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- DHFR- cells. Conclusion: A large amount of GPI modified Met-RANTES fusion protein was expressed on CHO cells. GPI anchored form of Met-RANTES may be used as novel immunosuppressant for suppressing reaction in graft rejection.
