A preliminary study on the expression and biological function of recombinant human SCF-TPO fusion protein / 中华血液学杂志
Chinese Journal of Hematology
;
(12): 19-22, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-229891
ABSTRACT
<p><b>OBJECTIVE</b>To study the expression of recombinant human SCF-TPO fusion protein and its biological function.</p><p><b>METHODS</b>Four primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography.</p><p><b>RESULTS</b>The high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml.</p><p><b>CONCLUSIONS</b>Expressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Physiology
/
Thrombopoietin
/
Recombinant Fusion Proteins
/
Cell Survival
/
Blotting, Western
/
Cloning, Molecular
/
Stem Cell Factor
/
Reverse Transcriptase Polymerase Chain Reaction
/
Cell Line, Tumor
/
Cell Proliferation
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Hematology
Year:
2005
Type:
Article
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