Expression in Escherichia coli and purification of human thrombopoietin.
Human thrombopoietin (TPO) has been successfully overexpressed in Escherichia coli, with an expression level of about 12% of total cellular protein. The full-length TPO gene was subcloned into the prokaryotic expression vector pKK233-2 under the control of the inducible tac promoter. The recombinant protein was produced mainly in the form of inclusion body. By efficient renaturation and one-step purification, the recombinant protein was purified to homogeneity. The specific activity and yield of recombinant TPO can reach 2 x 10(4) units/mg and 2 mg/g of wet E. coli cells respectively.
Li
S
S
Huadong Research Institute for Medical Biotechnics, Nanjing, People's Republic of China.
Li
F
F
Wu
W
W
Tan
W
W
Yu
M
M
Chen
J
J
Tao
K
K
D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013926:Thrombopoietin / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification
D011994:Recombinant Proteins; D013926:Thrombopoietin