This study was conducted to construct parasin Ⅰ expression vector, produce the recombinant peptide in the E.coli expression system, and analyze the antibacterial activity of the recombinant parasin Ⅰ. According to the amino acid sequence of mature peptide of parasin Ⅰ and codon bias of E. coli, a 57 bp cDNA fragment was synthesized. The recombinant vector pET32-Para Ⅰ in E. coli was constructed by PCR, and a sequence encoding endonuclease site of Xa factor was cloned into the 5'-upstream of the parasin Ⅰ gene. The recombinant vector was delivered into E. coli Rosetta (DE3) cells and isopropyl β-D-thiogalactoside (IPTG) was used to induce recombinant fusion protein under different temperatures (37 and 20 ℃). The results showed that the recombinant fusion protein reached about 45% to 50% of the total protein in E. coli, which had a molecular mass of 21 ku. The fusion protein induced under low temperature (20 ℃) mainly existed in dissolvable form. After purified by affinity chromatography, the recombinant parasin Ⅰ showed potent anti-Staphylococcus aureus activity after digested by Xa fator. An efficient expression system in E. coli for the production of a recombinant parasin Ⅰ with high yield are successfully constructed in this study, and the recombinant protein shows potent anti-bacterial activity after digested by Xa fator.
ZHAO Hua
,
ZHANG Yanyan
,
TANG Jiayong
,
LI Ke
,
ZHOU Jichang
,
WANG Kangning
. Parasin Ⅰ: Prokaryotic Expression, Purification and Antimicrobial Activity Analysis of a Recombination Catfish Antimicrobial Peptide[J]. Chinese Journal of Animal Nutrition, 2012
, 24(9)
: 1731
-1736
.
DOI: 10.3969/j.issn.1006-267x.2012.09.016
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