Prokaryotic expression and purification of His-tagged-STAT4 (565-748 amino acid) fusion protein / 中国免疫学杂志
Chinese Journal of Immunology
;
(12): 708-711,717, 2018.
Article
in Chinese
| WPRIM
| ID: wpr-702802
ABSTRACT
Objective:
To construct His-tagged peptide of human STAT4 (565-748 amino acid) expression vector and induce its expression in Escherichia coli,followed by purification.Methods:
STAT4 gene fragment encoding C-terminal peptide of 565-748 amino acid was amplified by PCR using pEGFP-STAT4 as the template.The PCR product was inserted into prokaryotic expression vector pET-28a and was transformed into component E.coli BL21 cells.By isopropyl-β-D-thiogalactoside(IPTG) induction,fusion protein was found to be expressed in the inclusion body and was denatured by using the urea denaturation buffer followed by renaturation and purifi-cation.Finally the purified protein was confirmed by Western blot.Results:
The STAT4 truncated gene encoding 565-748 amino acids peptide was amplified by PCR and inserted into pET-28a vector.After the recombined plasmid was transformed into component BL21, the His-tagged-STAT4 (565-748 amino acids) fusion protein was induced and obtained after denaturation,refolding,purification and dialysis.Conclusion:
The eukaryotic expression vector containing the truncated human STAT4 gene encoding 565-748aa peptide has been successfully constructed and the fusion protein was obtained.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Immunology
Year:
2018
Type:
Article
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