Molecular cloning and expression of a C‐type lectin‐like protein from orange‐spotted grouper Epinephelus coioides. Issue 2 (February 2014)

• Record Type: Journal Article
• Authors: Ji, H.; Wei, J.; Wei, S.; Yan, Y.; Huang, Y.; Huang, X.; Zhou, S.; Zhou, Y.; Qin, Q.
• Is Part Of: Journal of fish biology. Volume 84:Issue 2(2014)
• Page Start: 436
• Page End: 447
• Publication Date: 2014-02
• Subjects: Fishes -- Periodicals; Fishes -- Great Britain -- Periodicals; 597
• DOI: 10.1111/jfb.12296
• Languages: English
• ISSNs: 0022-1112

Abstract

<abstract abstract-type="main" id="jfb12296-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <p id="jfb12296-para-0001">A C‐type lectin‐like protein (Ec‐CTLP) was cloned from the grouper <italic>Epinephelus coioides</italic>. The full‐length cDNA of Ec‐CTLP was composed of 905 bp with a 522 bp open reading frame that encodes a 174‐residue protein. The putative amino acid sequence of Ec‐CTLP contains a signal peptide of 19 residues at the N‐terminus and a CLECT domain from Cys43 to Arg169 and a conserved imperfect WND (Trp‐Asn‐Asp) motif. The homologous identity of deduced amino acid sequences is from 32 to 42% with other fishes. The expression of Ec‐CTLP was differently upregulated in <italic>E. coioides</italic> spleen (germline stem) cells after being challenged at 16 and 4° C. Intracellular localization revealed that Ec‐CTLP was distributed only in the cytoplasm. Recombinant Ec‐CTLP (rEc‐CTLP) was expressed in <italic>Escherichia coli</italic> BL21 (DE3) and purified for mouse <italic>Mus musculus</italic> anti‐Ec‐CTLP serum preparation. The rEc‐CTLP fusion protein does not possess haemagglutinating activity, but improves survival from frozen bacteria. The survival of bacteria (including gram‐negative <italic>E. coli</italic> and gram‐positive <italic>Staphylococcus aureus</italic>) was positively correlated with the concentration of the rEc‐CTLP. These findings can provide clues to help understand the probable C‐type lectin in marine fish innate immunity.</p> </abstract>