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Apoptosis Induction and Detection in a Primary Culture of Sea Cucumber Intestinal Cells
Published on: January 21, 2020
Molecular characterization and functional analysis of MyD88 from the tropical sea cucumber, Holothuria leucospilota
Lin Zhao1, Xiao Jiang2, Ting Chen2
1Guangdong Provincial Key Laboratory of Biotechnology Candidate Drug Research, School of Biosciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou, 510006, PR China.
Abstract:
In this study, a myeloid differentiation factor 88 (MyD88) named as HLMyD88 was identified from the sea cucumber Holothuria leucospilota. The full-length cDNA of HLMyD88 is 4797 bp in size, containing a 227 bp 5'-untranslated region (UTR), a 3721 bp 3'-UTR and an 849 bp open reading frame (ORF) encoding a protein of 282 amino acids with a deduced molecular weight of 32.25 kDa HLMyD88 contains an N-terminal death domain and a C-terminal Toll/interluekin-1 receptor (TIR) domain with three highly conserved sequence motifs named as Box 1, Box 2 and Box 3. The results of luciferase reporter assay showed that over-expressed HLMyD88 in HEK293T cells could activate the transcription factors nuclear factor-κB (NF-κB) and activator protein 1 (AP-1). Additionally, the secretion of proinflammatory cytokines IL-1β and TNF-α in the HEK293T cells was increased by over-expressed HLMyD88, indicating the potential role of HLMyD88 in the innate immunity of sea cucumber. Moreover, we further confirmed that over-expressed HLMyD88 could also induce apoptosis.
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