Soluble expression and purification of human alpha-defensin-5 in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 291-296, 2008.
Artigo
em Chinês
| WPRIM
| ID: wpr-276125
ABSTRACT
DNA fragment containing human alpha-defensin 5 mature peptide (mHD-5) coding sequence with biased codons of E. coli was amplified by PCR, which was subsequently cloned into the plasmid pMAL-p2x in order to create pMAL-p2x-mHD-5 expression vector. The plasmid pMAL-p2x-mHD-5 was transferred into engineered strain BL21(DE3) to express heterogeneous fusion protein (MBP-mHD-5). The soluble MBP-mHD-5 targeted protein inducible expressed by IPTG was accounted for about 30% under optimized conditions. The recombinant mHD-5 (rmHD-5) peptide was successfully purified through a separation process including affinity chromatography, Factor Xa digestion and ion exchange chromatography. The bioactivity of rmHD-5 was examined by bacteria-inhibition tests in liquid culture. The growth of E. coli ATCC25922 was dramatically suppressed with an inhibition rate of 90%, with the presence of 62.5 microg/mL rmHD-5 in the media. These results indicate that the strategy of soluble expression of fusion protein in E. coli can be a useful and practical way to produce bioactive defensins.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Solubilidade
/
Transformação Bacteriana
/
Proteínas Recombinantes de Fusão
/
Clonagem Molecular
/
Alfa-Defensinas
/
Escherichia coli
/
Genética
/
Metabolismo
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2008
Tipo de documento:
Artigo
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