Prokaryotic expression, purification, and identification of recombinant human IL-11 protein.
- Author:
Jia TANG
1
;
Xiaoling XU
;
Xiaoxia NIE
;
Qifeng MAO
;
Jimin GAO
Author Information
1. Zhejiang Provincial Key Lab for Model Organisms, Wenzhou Medical College, Wenzhou 325035, China.
- Publication Type:Journal Article
- MeSH:
Escherichia coli;
metabolism;
Genetic Vectors;
genetics;
Humans;
Interleukin-11;
genetics;
metabolism;
Recombinant Proteins;
genetics;
isolation & purification;
metabolism
- From:
Journal of Biomedical Engineering
2012;29(3):530-533
- CountryChina
- Language:Chinese
-
Abstract:
A DNA fragment encoding recombinant human interleukin 11 (hrIL-11) was obtained by PCR from previously constructed pET24a-hrIL-11 plasmid. Then pET21a-hrIL-11 expression vector was constructed routinely and transformed into BL-21(DE3). By the induction of Isopropyl-1-thio-beta-D-galactoside (IPTG), hrIL-11 protein was highly expressed at about 20% of the total bacterial proteins and was identified by Western blot. After purification with Ni-NTA affinity chromatography and refolding with renaturation buffer, the purity of the target hrIL-11 protein reached 95% and its biology activity was 1 x 10(6) IU/mg, determined by stimulating the proliferation of T1165, which facilitates further researches into effects of IL-11 on platelet proliferation and other function.
