[Construction, expression and biologic activities of two rhIL-6/GM-CSF fusion proteins].
OBJECTIVE
To construct and express the gene encoding hIL-6/GM-CSF fusion protein.


METHODS
The genes encoding the two hIL-6/GM-CSF fusion proteins were constructed in pBV220 expression vector. Fusion proteins were expressed in E.coli BL-21 and purified through Q Sepharose HP ion exchange chromatography and Sephacryl S-200 gel filtration columns. The biologic activities of the fusion proteins were detected by proliferation of hIL-6 dependent cell line B9 and hGM-CSF dependent cell line TF1 with MTT assay.


RESULTS
Both fusion proteins were expressed in E.coli BL-21 in the form of inclusion body. The expression levels were more than 25% of the total cell lysates. Both fusion proteins were obtained with high purity which had both hIL-6 and hGM-CSF biologic activities.


CONCLUSIONS
Two hIL-6/GM-CSF fusion proteins with high purity and bilolgic activities have been acquired successfully.
Sun
Qiang-ming
QM
Department of Molecular Biology, Institute of Medical Biology, Chinese Academy of Medical Sciences, Peking Union Medical College, Kunming 650118, China. sunqiangming1@sina.com
Sun
Mao-sheng
MS
Yi
Shan
S
Cao
Zeng
Z
Bie
Jun
J
Dai
Chang-bai
CB
Xu
Wei-ming
WM
D015153:Blotting, Western; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics* / Q000494:pharmacology; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics* / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology
D015850:Interleukin-6; D011993:Recombinant Fusion Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor