[Construction of an expression vector with elastin-like polypeptide tag to purify xylanase].
Wei Sheng Wu Xue Bao
; 52(1): 90-5, 2012 Jan.
Article
en Zh
| MEDLINE
| ID: mdl-22489465
ABSTRACT
OBJECTIVE:
This paper reports the purification of xylanase using the shortest, sensitive ELP [KV8F-20].METHODS:
We designed a thermophilic xylanase gene, and recombined it with the ELP via a random coil sequence to generate the vector pET-22b-SoxB-M2-S-ELP. The expressed xylanase was purified by inverse transition cycling through high-speed centrifugation, and then we characterized the purified xylanase.RESULTS:
The phase transition temperature of the ELPs dropped to 22 degrees C with 0.5 mol/L sodium carbonate (pH = 7). Under this condition, SoxB-M2-S-ELP was purified by 3.16 folds after centrifugation. The recovery rate was 21.2%, and purity of the xylanase was 64.3%.CONCLUSION:
Elastin-like polypeptide as a purification tag to purify recombinant proteins is simple, fast, gentle and cheaper. The expression vector we constructed here might be a very useful and reliable tool to purify many other target proteins.
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Banco de datos:
MEDLINE
Asunto principal:
Péptidos
/
Elastina
/
Endo-1,4-beta Xilanasas
/
Vectores Genéticos
Idioma:
Zh
Año:
2012
Tipo del documento:
Article