›› 2013, Vol. 25 ›› Issue (6): 0-1214.
• 动物科学 •
WU Jing;LIANG Yong\|li;SHI Yyu\|ying;LI Yyu\|feng;MA Xiu\|li;JIANG Yi\|fei;SONG Min\|xun;*
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Abstract: Cathelicidins comprise a family of antimicrobial peptides sharing a highly conserved cathelin domain, they play a critical role in the innate immune system of chicken. In order to investigate the antibacterial activity of recombinant Cathelicidins, the putative mature peptide coding fragments of CathL\|1, CathL\|2, CathL\|3 genes were amplified from the plasmid pMD\|CathL containing Cathelicidins cDNA sequences, and then subcloned into pET\|30a(+) plasmid respectively. The recombinant vectors named pET30\|CathL1S, pET30\|CathL2S and pET30\|CathL3S were transformed into the competent cell of E.coli Rosetta(DE3). The positive clones were cultured and induced to express target protein by addition of 10 mmol·L-1 IPTG in LB. SDS\|PAGE and Western blot analysis demonstrated that cathelicidins genes were expressed in the form of fusion protein, and the apparent molecular weight of expression products were 70 kD, 80 kD, 75 kD, respectively. The recombinant Cathelicidins were able to kill both Gram negative bacteria and Gram positive bacteria by the means of agar well diffusion assay (AWDA), especially for the antibiotic\|resistant strains.
Key words: Silkie, Cathelicidins antimicrobial peptide, fusion expression, purification, antibacterial activity
WU Jing;LIANG Yong\|li;SHI Yyu\|ying;LI Yyu\|feng;MA Xiu\|li;JIANG Yi\|fei;SONG Min\|xun;* . Fusion expression of Silkie Cathelicidins genes in Escherichia coli and detection of their antibacterial activity[J]. , 2013, 25(6): 0-1214.
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