Efficient fusion expression of G13 domain derived from granulysin in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 235-241, 2009.
Article
in Chinese
| WPRIM
| ID: wpr-302830
ABSTRACT
The G13 domain derived from granulysin shows high antimicrobial activities against Gram-positive and Gram-negative bacteria but does not lyse Jurkat cells or liposomes. To explore a new approach for high expression of the G13 domain, we fused the sequence encoding G13 to thioredoxin (Trx) gene to construct the recombinant expression vector (pThioHisA-G13). A cyanogen bromide (CNBr) cleavage site was introduced between the Trx and G13 to facilitate final release of the recombinant G13. The recombinant expression vector, pThioHisA-G13, was transformed into E. coli BL21 (DE3). Upon induction by IPTG Trx-G13 fusion protein was expressed and took the form of inclusion bodies counting 58% (W/W) of total cellular proteins. The inclusion body was solved by urea (8 mol/L) and then cleaved by CNBr. We purified the recombinant peptide G13 by one-step cation exchange chromatography. Results of agarose diffuse assay analysis indicated that the recombinant G13 exhibited antibacterial activity. The procedure described in this study will provide a reliable and simple method for highly efficient production of some cationic antimicrobial peptides.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Thioredoxins
/
Recombinant Fusion Proteins
/
Transfection
/
Antigens, Differentiation, T-Lymphocyte
/
Inclusion Bodies
/
Protein Structure, Tertiary
/
Cyanogen Bromide
/
GTP-Binding Protein alpha Subunits, G12-G13
/
Escherichia coli
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2009
Type:
Article
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