Main content area

A new leukocyte cell-derived chemotaxin-2 from marine fish grouper, Epinephelus coioides: Molecular cloning and expression analysis

Wei, Jingguang, Guo, Minglan, Cui, Huachun, Yan, Yang, Ouyang, Zhengliang, Qin, Qiwei
Fish & shellfish immunology 2011 v.31 no.4 pp. 600-605
Epinephelus coioides, Escherichia coli, Larimichthys crocea, Saccharomyces cerevisiae, Singapore grouper iridovirus, Staphylococcus aureus, Vibrio parahaemolyticus, Vibrio vulnificus, antiserum, autoimmunity, cell growth, complementary DNA, concanavalin A, freshwater fish, genes, grouper, innate immunity, leukocytes, liver, marine fish, messenger RNA, molecular cloning, open reading frames, rapid amplification of cDNA ends
Leukocyte cell-derived chemotaxin-2 (LECT2) is a multifunctional protein involved in cell growth, differentiation and autoimmunity. In this study, a new leukocyte cell-derived chemotaxin-2 (EcLECT2) gene was cloned from grouper, Epinephelus coioides, by rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA sequence of EcLECT2 was 595 bp in size, containing a 5′-untranslated region (UTR) of 44 bp and a 3′-UTR of 83 bp. The deduced protein sequence of the open reading frame (465 bp) showed highest similarity (81%) to the LECT2 of the fresh-water fish Larimichthys crocea. An abundant transcription of the determined EcLECT2 mRNA has been detected in liver and skin of grouper, E. coioides. Furthermore, the expression of EcLECT2 was differentially up-regulated in liver after infection with Staphyloccocus aureus, Vibrio vulnificus, Vibrio parahaemolyticus, Saccharomyces cerevisiae and Singapore grouper iridovirus (SGIV), while the expression was down-regulated after stimulation with Concanavalin A (Con A). Recombinant mature EcLECT2 (rEcLECT2) was successfully expressed in Escherichia coli BL21 (DE3), and the antiserum against EcLECT2 was obtained for further investigations. EcLECT2 may be an important molecule in the innate immunity of grouper.