Molecular cloning and expression of MyD88 in large yellow croaker, Pseudosciaena crocea.
Myeloid differentiation factor 88 (MyD88) is an adaptor protein involved in the interleukin-1 receptor and Toll-like receptor-induced activation of nuclear factor-kappaB (NF-kappaB). In this report, the full-length cDNA of MyD88 was cloned from the large yellow croaker, Pseudosciaena crocea. It was of 1574 bp, including a 5'-terminal untranslated region (UTR) of 89 bp, a 3'-terminal UTR of 621bp and an open reading frame (ORF) of 864 bp encoding a polypeptide of 287 amino acids. It contained a typical death domain at the N-terminal and a conservative Toll/IL-1R (TIR) domain structure at the C-terminal. The quantitative real-time reverse transcription PCR analysis revealed a broad expression of MyD88 with the highest expression in the spleen and the weakest expression in the muscle. The expression of MyD88 after challenge with formalin-inactivated Gram-negative bacterium Vibrio parahaemolyticus was tested in blood, spleen and liver. It suggested that the highest expression was in the spleen (p<0.05) with 1.9 times (at 48 h) as much as that in the control and the lowest expression of MyD88 was in the liver (p<0.05) with 0.29 times (at 3h) of that in the control. These results indicated that as a universal key adaptor in the Toll-like receptor pathway in mammals, MyD88 might play an important role in large yellow croaker defense against pathogenic infection.
Yao
Cui-Luan
CL
Key Laboratory of Science and Technology for Aquaculture and Food Safety of Fujian Province University, Fisheries College/Fisheries Biotechnology Institute, Jimei University, Xiamen 361021, China.
Kong
Peng
P
Wang
Zhi-Yong
ZY
Ji
Pei-Feng
PF
Liu
Xian-De
XD
Cai
Ming-Yi
MY
Han
Xue-Zhe
XZ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D008099:Liver / Q000276:immunology; D008969:Molecular Sequence Data; D053594:Myeloid Differentiation Factor 88 / Q000097:blood / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D010473:Perciformes / Q000145:classification / Q000235:genetics* / Q000276:immunology*; D010802:Phylogeny; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000276:immunology; D014735:Vibrio Infections / Q000276:immunology / Q000662:veterinary; D014736:Vibrio parahaemolyticus / Q000276:immunology
D053594:Myeloid Differentiation Factor 88