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Molecular cloning and characterization of a C‐type lectin in yellow catfish Tachysurus fulvidraco
Author(s) -
Ke F.,
Zhang H. B.,
Wang Y.,
Hou L. F.,
Dong H. J.,
Wang Z. F.,
Pan G. W.,
Cao X. Y.
Publication year - 2016
Publication title -
journal of fish biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.672
H-Index - 115
eISSN - 1095-8649
pISSN - 0022-1112
DOI - 10.1111/jfb.13080
Subject(s) - biology , c type lectin , catfish , lectin , microbiology and biotechnology , ictalurus , peptide sequence , open reading frame , ctl* , molecular cloning , recombinant dna , biochemistry , gene , cytotoxic t cell , fishery , fish <actinopterygii> , in vitro
This study represents the first report of a C‐type lectin ( ctl ) in yellow catfish Tachysurus fulvidraco . The complete sequence of ctl complementary (c) DNA consisted of 685 nucleotides. The open reading frame potentially encoded a protein of 177 amino acids with a calculated molecular mass of c. y 20.204 kDa . The deduced amino‐acid sequence contained a signal peptide and a single carbohydrate recognition domain with four cysteine residues and GlnProAsp ( QPD ) and TrpAsnAsp ( WND ) motifs. Ctl showed the highest identity (56.0%) to the predicted lactose binding lectin from channel catfish Ictalurus punctatus . Quantitative real‐time (qrt)‐ PCR analysis showed that ctl messenger (m) RNA was constitutively expressed in all examined tissues in normal fish, with high expression in trunk kidney and head kidney, which was increased following Aeromonas hydrophila challenge in a duration‐dependent manner. Purified recombinant Ctl ( rCtl ) from Escherichia coli BL21 was able to bind and agglutinate Gram‐positive and Gram‐negative bacteria in a calcium‐dependent manner. These results suggested that Ctl might be a C‐type lectin of T. fulvidraco involved in innate immune responses as receptors ( PRR ).