Construction, expression and activity evaluation of recombinant NuBCP-9/ Tumstatin(74-98) fusion polypeptide / 中国药科大学学报
Journal of China Pharmaceutical University
;
(6): 364-369, 2009.
Artigo
em Chinês
| WPRIM
| ID: wpr-480388
ABSTRACT
Aim:
To construct a prokaryotic expression vector carrying NuBCP-9-tumstatin(74-98) (abbreviated as NT) gene and to obtain the fusion peptide with antitumor activity.Methods:
Nucleotide sequences of antitumor peptides, NuBCP-9 and Tumstatin( 74-98), were connected via a linker(G_4S)_3 based on biased codons of E. coli the fused NT gene was reconstructed using SOE PCR, and inserted into pET32a(+) vector, and transformed in E. coli BL21(DE3). After expression, the novel fusion peptide was purified through nickel-affinity chromatogra-phy, Factor Xa digestion and ultrafiltration. Biological activity of the fusion peptide on ECV304 and A549 cells was evaluated by MTT assay.Results:
A prokaryotic expression system with NT gene was successfully constructed. The soluble fusion peptide was accounted for approximately 25% when induced by 0. 5 mmol/L IPTG at 30 ℃ for 4 h. The purified fusion peptide could inhibit cell growth of ECV304 and A549 with inhibition rates of 60. 8% and 65. 2% at 20 μmol/L, respectively.Conclusion:
A novel fusion peptide with antitumor activity was cloned, expressed and purified.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Journal of China Pharmaceutical University
Ano de publicação:
2009
Tipo de documento:
Artigo
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