Efficient expression and primary purification of 6-his tagged human Fas ligand in Dictyostelium discoideum.
Human Fas ligand (hFasL) is a member of the tumor necrosis factor (TNF) family with many medical interests. To produce this protein efficiently, an improved vector which could express the recombinant hFasL protein with a 6-his tag at its C-terminal was constructed. The new vector was transformed into Dictyostelium discoideum AX3 which then produced 157 microg hFasL l(-1). Using one-step Ni-affinity chromatography, it was purified with a recovery of 92% and purity of 91%.
Chen
Jie
J
Department of Chemical Engineering and Bioengineering, Zhejiang University, Hangzhou, PR China.
Lu
Yinghua
Y
Xu
Zhinan
Z
Cen
Peilin
P
Fang
Xiangming
X
D000818:Animals; D004023:Dictyostelium / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D053222:Fas Ligand Protein / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism*
D053222:Fas Ligand Protein; D011994:Recombinant Proteins