Cloning and Prokaryotic Expression of IL-10 Gene of Epinephelus coioides

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.018

Abstract

Abstract:

Interleukin 10(IL-10)is the critical multiple-functional cytokine playing the major role of mediating inflammatory responses and regulating the differentiation and proliferation of some immune cells. The full-length cDNA of IL-10 of Epinephelus coioides was cloned using homological cloning and rapid amplification of cDNA ends(RACE-PCR). Results showed the full-length of IL-10's cDNA was 1 104 bp, containing an open reading frame of 564 bp encoding 187 amino acids with an estimated molecular weight of 21.7 kD and an estimated isoelectric point of 5.74, and it had a signal peptide of 22 amino acid residues and no transmembranous region. Amino acid homology analysis showed that the amino acid sequences of IL-10 with Oreochromis niloticus had the highest homology and up to 72.25%. SDS-PAGE indicated that the molecular weight of the fusion protein was 37.5 kD which was in accord with the estimated. The optimal condition of inducible expression for IL-10 protein in Escherichia coli was at 37℃ for 3 h with 0.02 mmol/L of IPTG.

Key words: Epinephelus coioides, Interleukin-10, gene cloning, prokaryotic expression

Xiao Zhouting, Huang Yucong, Jian Jichang, Lu Yishan, Wu Zaohe,. Cloning and Prokaryotic Expression of IL-10 Gene of Epinephelus coioides[J]. Biotechnology Bulletin, 2015, 31(9): 131-137.

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