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Molecular cloning and expression analysis of myd88 from oriental weatherfish (Misgurnus anguillicaudatus) in response to bacterial challenge

JOURNAL OF FISH BIOLOGY(2020)

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Abstract
Myeloid differentiation factor 88 (Myd88) plays an important role in both innate and adaptive immune response. In this study, the full-length complementary DNA (cDNA) ofmyd88fromMisgurnus anguillicaudatuswas characterized. Themyd88cDNA is 1333 bp in length and contains an 855 bp open reading frame encoding a predicted protein of 284 amino acids. The predicted protein possesses typical Myd88 domain structural features including a death domain in the N-terminus, and box 1, 2, and 3 motifs of the Toll/IL-1 receptor domain in the C-terminus. Quantitative real-time PCR (qRT-PCR) revealed thatmyd88messenger RNA (mRNA) was ubiquitously expressed in all examined tissues, especially highly in brain, kidney, blood, intestines and liver. qRT-PCR and western blotting were used to determine the mRNA and protein levels of Myd88 afterAeromonas veroniichallenge, respectively. The Myd88 was remarkably upregulated in response to infection ofA. veronii.These results suggested that Myd88 may play a vital role during the immune response ofM. anguillicaudatusagainst bacterial infection.
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Key words
bacterial challenge,expression analysis,Misgurnus anguillicaudatus,molecular characterization,myd88
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