Recombinant production of anti-HIV protein, griffithsin, by auto-induction in a fermentor culture》.pdf

Protein Expression and PuriWcation 47 (2006) 194–202 /locate/yprep Recombinant production of anti-HIV protein, griYthsin, by auto-induction in a fermentor culture Barbara Giomarelli a, Kathryn M. Schumacher a,b, Troy E. Taylor c, Raymond C. Sowder II d, James L. Hartley c, James B. McMahon a, Toshiyuki Mori a,¤,1 a Molecular Targets Development Program, Center for Cancer Research, National Cancer Institute, NCI-Frederick, Frederick, MD 21702-1201, USA b Werner H. Kirsten Student Internship Program, Center for Cancer Research, National Cancer Institute, NCI-Frederick, Frederick, MD 21702-1201, USA c Protein Expression Laboratory, Research Technology Program, SAIC-Frederick, Inc., Frederick, MD 21702, USA d AIDS Vaccine Program, SAIC-Frederick, Inc., Frederick, MD 21702, USA Received 18 August 2005, and in revised form 11 October 2005 Available online 7 November 2005 Abstract GriYthsin (GRFT) is a novel anti-HIV protein isolated from the red alga GriY thia sp. The potent anti-viral activity of GRFT against both laboratory and primary isolates of HIV at picomolar concentrations makes this protein an attractive candidate microbicide to pre- vent sexual transmission of HIV. Here, we describe the recombinant production and puriWcation of a biologically active hexa-histidine- tagged GRFT (His-GRFT) from Escherichia coli. To facilitate a large-scale production of recombinant His-GRFT, we tested diVerent expression conditions to optimize the expression in the cytoplasm of E. coli to increase the overall production of solub