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Identification, expression pattern and functional characterization of As-MyD88 in bacteria challenge and during different developmental stages of Artemia sinica

Qin, Tong, Zhao, Xinxin, Luan, Hong, Ba, Huazhong, Yang, Lei, Li, Zhenegmin, Hou, Lin, Zou, Xiangyang
Developmental and comparative immunology 2015 v.50 no.1 pp. 9-18
Artemia sinica, Toll-like receptors, Western blotting, amino acids, bacteria, complementary DNA, developmental stages, embryogenesis, immune response, immunohistochemistry, open reading frames, quantitative polymerase chain reaction
Myeloid differentiation factor 88 (MYD88), a key adapter protein in Toll-like receptor signaling, affects the immune response and the formation of the dorsal–ventral axis. Here, the 1555bp full-length cDNA of MyD88 from Artemia sinica (As-MyD88) was obtained. Molecular characterization revealed that the sequence includes an 1182bp open reading frame encoding a predicted protein of 393 amino acids. The predicted protein contains a death domain in the N-terminus, and box1 and 2 motifs of the TIR domain in the C-terminus. Real-time quantitative PCR, Western blotting and immunohistochemistry were used to determine the expression level, protein production and location of As-MYD88 during embryonic development and bacterial challenge. The highest expression level during embryonic development was at the 0h and 5h stages of A. sinica. As-MYD88 was remarkably upregulated after bacterial challenge. Our results suggested that As-MYD88 plays a vital role in response to bacterial challenge, and during post-diapause embryonic development of A. sinica.