Optimization of Expression of Recombinant Human Tumor Necrosis Factor(hTNF-α) in Escherichia coliE.coli BL21 (DE3) / 现代检验医学杂志
Journal of Modern Laboratory Medicine
;
(4): 100-103,107, 2017.
Artículo
en Chino
| WPRIM
| ID: wpr-667246
ABSTRACT
Objective To construct a human tumor necrosis factor (hTNF-a) plasmid and identify it to optimize the fermentation conditions of hTNF-α protein so as to achieve high expression in Escherichia coli.Methods The gene of hTNF-a was cloned into pET24a vector to obtain the pET24a-hTNF-a expression plasmid that was transformed into Escherichia coli BL21(DE3),and the expression conditions of BL21 (DE3) were optimized.Results The plasmid of pET24a-hTNF-α was successfully constructed and identified by PCR and digestion,which was consistent with the target fragment hTNF-α.The plasmid was transformed into Escherichia coli BL21(DE3),the best induced expression conditions of Escherichia coli BL21 (DE3) were as followsM9+LB medium,37℃,0.5 mmol/L IPTG,pH =7.5,and induction time was 5 h.The results showed that dry weight of the cells and the rate of TNF were increased by 2.56 times and 3.68 times,respectively,and the expression rate of hTNF-α was increased by 3.49 times from 9.38% to 32.74%.Conclusion The optimal conditions for the expression of plasmid pET24a-hTNF-α in Escherichia coli were determined.
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Índice:
WPRIM (Pacífico Occidental)
Tipo de estudio:
Estudio pronóstico
Idioma:
Chino
Revista:
Journal of Modern Laboratory Medicine
Año:
2017
Tipo del documento:
Artículo
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