Expression of Tumstatin183-230-TRAIL fusion protein and identification of its biological functions / 第二军医大学学报
Academic Journal of Second Military Medical University
;
(12): 474-478, 2010.
Artigo
em Chinês
| WPRIM
| ID: wpr-840867
ABSTRACT
Objective:
To express Tumstatin183-230-TRAIL fusion protein and to observe its biological functions.Methods:
SOE-ing PCR was employed to amplify the recombinant sequence of Tumstatin183-230 and TNF-related apoptosis-inducing ligand (TRAIL114-281). An expression vector pMAL-Tu-T was constructed by inserting Tu-T sequence into pMAL-c2; the vector was used to transfect E. coli BL21 (DE3) and expression of MBP-Tu-T fusion protein was induced by IPTG. Amylose Resin columns were employed to purify the fusion protein. The biological functions of MBP-Tu-T protein was examined by inhibitory test of endothelial cell proliferation, standard tumor cell cytotoxic assay, in vitro tube formation inhibition, and electron microscopic observation (apoptosis).Results:
The expression rate of MBP-Tu-T fusion protein in E. coli was about 20%. Purified recombinant protein obviously inhibited endothelial cell proliferation (IC50 12.5 μg/ml), induced apoptosis of pancreatic cancer cells, and inhibited tube formation.Conclusion:
Constructed MBP-Tu-T fusion protein is bifunctional, which lays a solid foundation for further investigation of antitumor effect of Tumstatin183-230-TRAIL in vivo.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Academic Journal of Second Military Medical University
Ano de publicação:
2010
Tipo de documento:
Artigo
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