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    STUDIA CHEMIA - Issue no. Special Issue / 2010  
         
  Article:   PURIFICATION AND CLEAVING OF THE HUMAN GITRL EXPRESSED AS FUSION PROTEIN.

Authors:  . 		 
       
         
  Abstract:  The glucocorticoid-induced tumor necrosis factor (TNF) receptor (GITR) is a member of the TNF receptor superfamily. GITR is activated by its ligand, GITRL. GITRL is a type II transmembrane protein. Human tumor cells express high levels of the GITRL; the presence of it thus can be used as a potential tumor marker. In the present study, the constructed pETM52-hGITRL recombinant vector was transformed into the chemically competent Escherichia coli BL21 Star (DE3) strain. After induction with 0.5 mM isopropyl β-D-thiogalactoside (IPTG), the expression was maintained at 37oC for an additional 4h. The protein was obtained in sufficient quantity for refolding studies, after which it was purified by affinity chromatography then cleaved from the fusion partner by tobacco etch virus (TEV) protease treatment.

Keywords: GITR, human GITRL, tumor marker, affinity chromatography, TEV protease 

  
         
     
         
         
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