Molecular characterization of Fyn, Lck and Zap-70 in golden pompano (Trachinotus ovatus).
TL;DR
Co-immunoprecipitation assay demonstrated that both trFyn and trLck could interact with trZap-70 in HEK-293T cells, providing molecular basis for further study of the TCR signaling and T-cell mediated immunity in golden pompano.
Abstract
T-cell receptor (TCR) signaling is essential for adaptive immunity in vertebrates, with the tyrosine kinases Fyn, Lck, and Zap-70 playing central roles in signal transduction. In the present study, the full cDNA sequences of Fyn, Lck, and Zap-70 from golden pompano (Trachinotus ovatus) were obtained by RT-PCR and RACE-PCR and their expression patterns in normal tissues and head kidney (HK) following lipopolysaccharide (LPS), polyinosinic-polycytidylic acid (polyI:C) stimulation and Vibrio alginolyticus infection were examined by qPCR. The open reading frames of trFyn, trLck, and trZap-70 respectively encoded 537, 513, and 609 amino acids, all containing conserved similar functional domains as their mammalian counterparts. qPCR analysis revealed that all three genes were ubiquitously expressed in examined tissues, with high expression in immune-related tissues. trFyn, trLck, and trZap-70 were significantly induced in HK after LPS, polyI:C stimulation and V. alginolyticus infection. Co-immunoprecipitation (co-IP) assay demonstrated that both trFyn and trLck could interact with trZap-70 in HEK-293T cells. These findings provide molecular basis for further study of the TCR signaling and T-cell mediated immunity in golden pompano.