Construction of prokaryotic expression vector and expression of human beta defensin 4 gene / 生物医学工程学杂志
Journal of Biomedical Engineering
;
(6): 1166-1169, 2008.
Article
in Chinese
| WPRIM
| ID: wpr-318192
ABSTRACT
Human beta defensin 4 is a small cationic peptide with a broad range of antimicrobial activity. It plays an important role in innate immunity of human body, especially in mucosal and epithelial defense. In this study, the full-length encoding gene of HBD4 was synthesized by overlap extension polymerase chain reaction and inserted into cloning vector pMD18-T. The gene encoding mature peptide of HBD4 was amplified by PCR and cloned into prokaryotic expression vector pGEX-4T-2. Then pGEX-4T-2/mHBD4 was transformed into E. coli DH5 alpha, which was induced by isopropy-beta-D-thiogalactoside (IPTG). The identification was made by means of endonuclease digestion, DNA sequencing, sodium dodecyl sulphate-polyacrylamine gel electrophoresis (SDS-PAGE). The results showed that the synthesized gene and cloned gene were identical to the HBD4 gene sequence registered in GenBank and were successfully cloned into cloning vector pMD18-T and prokaryotic expression vector pGEX-4T-2. After IPTG induction, the GST-HBD4 fusion protein was successfully expressed in E. coli.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombinant Fusion Proteins
/
Cloning, Molecular
/
Beta-Defensins
/
Electrophoresis, Polyacrylamide Gel
/
Escherichia coli
/
Genetic Vectors
/
Genetics
/
Metabolism
/
Anti-Infective Agents
Limits:
Humans
Language:
Chinese
Journal:
Journal of Biomedical Engineering
Year:
2008
Type:
Article
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