2005•Xumu shouyi xuebaoRequires access
Cloning of chicken interleukin--18 mature protein gene and its high expression in {\sl E. coli}
Jingdong Hu, Zhizhong Cui, Zhao HongKun
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Abstract
Chicken interleukin-18 (ChIL-18) mature protein gene was amplified from LPS-stimulated MDCC-MSB1 cells by RT-PCR.PCR product was cloned into the pMD18-T vector. Sequencing analysis showed that the nucleotide sequence of this ChIL-18 mature protein gene was 5l0bp including the stop coden and the same as the published ChIL-18 cDNA sequence by Schneider K. A prokaryotic expression plasmid of ChIL-18, pGEX-ChIL18, was obtained by subcloning the encoding region of the ChIL-18 mature peptide into pGEX-6P-1. The recombinant ChIL-18 (rChIL-18) was expressed efficiently in pGEX-ChIL18-transformed BL21(DE3)LysS induced by IPTG, the yield was accounted for 32% of the total bacterial protein.