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Development of consensus qPCR primers to detect cytokine genes in three amberjack species: Seriola quinqueradiata, S. lalandi and S. dumerili

Author:
Jirapongpairoj, Walissara, Kobayashi, Keigo, Fukuda, Yutaka, Takano, Tomokazu, Sakai, Takamitsu, Matsuyama, Tomomasa, Nakayasu, Chihaya, Nozaki, Reiko, Hirono, Ikuo, Kondo, Hidehiro
Source:
Fisheries science 2015 v.81 no.5 pp. 907-914
ISSN:
0919-9268
Subject:
Seriola quinqueradiata, Vibrio anguillarum, gene expression regulation, genes, high-throughput nucleotide sequencing, interleukin-10, interleukin-1beta, interleukin-8, kidneys, pathogens, polyinosinic-polycytidylic acid, quantitative polymerase chain reaction, tumor necrosis factor-alpha, vaccination
Abstract:
Partial sequences of cytokine genes IFNγ, IL-1β, IL-8, IL-10, and TNFα were identified from next-generation sequencing data for three amberjack species: Seriola quinqueradiata, S. lalandi and S. dumerili. The cytokine genes had more than 90 % nucleotide identity among the three species. Quantitative PCR (qPCR) primers were designed from the consensus sequences to detect cytokine genes. The specificities of consensus qPCR primers were investigated in the head kidney of three amberjack species treated with formalin-killed cells (FKC) of Vibrio anguillarum and polyinosinic-polycytidylic acid [poly(I:C)] at 3 h post injection (hpi) using qPCR. The primers detected up-regulation of certain cytokine genes after FKC of V. anguillarum and poly(I:C) stimulation in all species examined. Consequently, the primers are suitable for studying and detecting cytokine genes after vaccination and pathogen infection in these three amberjack species.
Agid:
4405726