Molecular cloning and expression of human granulocyte-macrophage colony stimulating factor in Escherichia coli system.
Human granulocyte-macrophage stimulating factor (hGM-CSF) is an important hematopoietic growth factor. hGM-CSF is able to stimulate hematopoietic progenitors to produce granulocytes, macrophages and eosinophils. hGM-CSF also shows the potential to treat myeloid leukemias. In order to prepare enough hGM-CSF, a genetically engineered strain E. coli HB101/pZW.GM47 was constructed by DNA recombination technologies in the present study, and characterized by restriction analysis, DNA sequencing, SDS-PAGE, Western blotting and bioassays. The strain has been shown to produce a 14kDa mature hGM-CSF protein with expression level higher than 40% and a specific activity of 5.0 x 10(7) u/mg which shows a good potential for clinical applications.
Wang
J
J
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing, China.
Zou
M
M
Huang
B
B
Zhang
M
M
Mao
N
N
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D002455:Cell Division / Q000235:genetics; D002478:Cells, Cultured / Q000166:cytology / Q000502:physiology; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics*; D005798:Genes, Bacterial; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D001426:Bacterial Proteins; D004274:DNA, Recombinant; D016178:Granulocyte-Macrophage Colony-Stimulating Factor