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
Dans Zh
| WPRIM
| ID: wpr-229891
Responsable en Bibliothèque :
WPRO
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>
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Physiologie
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Thrombopoïétine
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Protéines de fusion recombinantes
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Survie cellulaire
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Technique de Western
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Clonage moléculaire
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Facteur de croissance des cellules souches
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RT-PCR
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Lignée cellulaire tumorale
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Prolifération cellulaire
Limites du sujet:
Humans
langue:
Zh
Texte intégral:
Chinese Journal of Hematology
Année:
2005
Type:
Article