The lipopolysaccharide-induced TNF-alpha factor (LITAF) is an inflammatory cytokine, which plays an important role in innate immunity system. Based on the expressed sequence tag (EST) of Japanese scallop (Mizuhopecten yessoensis), the cDNA of LITAF gene was amplified using rapid amplification of cDNA ends (RACE) approach. Results showed that the full-length cDNA of LITAF is 1 551 bp consisting of a 5′ untranslated region (UTR) of 76 bp, a 3' UTR of 1 001 bp, and an open reading frame (ORF) of 474 bp encoding a polypeptide of 157 amino acids, and there is a conserved LITAF domain in amino acid sequences. The estimated molecular mass is 16.99 kDa and the theoretical isoelectric point is 6.24. The total length of LITAF is 3 698 bp, which includes three exons and two introns. Real-time quantitative PCR was carried out to measure LITAF mRNA expression in adult tissues and monitor mRNA expression patterns during embryonic development after bacteria (Vibrio anguillarum) challenged. The expression level of LITAF mRNA was detected in all the adult tissues with the highest in the kidneys. The trochophore owns the highest expression level of LITAF in embryonic development. LITAF expression showed significant difference(P<0.01)between the control and bacteria challenged specimens at 36 h. These results suggest that the LITAF should be a member of the LITAF family that perhaps involved in the innate immune response of Japanese scallop.
HE Chong-Bo, ZHU Bao, LIU Wei-Dong, BAO Xiang-Bo, LI Yun-Feng, SHAN Zhong-Guo, LI Hong-Jun
. Cloning and expression analysis of lipopolysaccharide-induced TNF-a(LITAF) of Japanese scallop (Mizuhopecten yessoensis)[J]. Hereditas(Beijing), 2012
, 34(6)
: 736
-741
.
DOI: 10.3724/SP.J.1005.2012.00736
[1] Loker ES, Adema CM, Zhang SM, Zhang SM, Kepler TB. Invertebrate immune systems-not homogeneous not simple, not well understood. Immunol Rev, 2004, 198(1): 10-24.
[2] Chappell VL, Le LX, LaGrone L, Mileski WJ. STAT proteins play a role in tumor necrosis factor alpha gene expression. Shock, 2000, 14(3): 400-403.
[3] Marrack P, Kappler J. T cells can distinguish between al-logeneic major histocompatibility complex products on different cell types. Nature, 1988, 332(6167): 840-843.
[4] 向萍霞, 李艳, 张平安. 冠心病患者TNF-α水平及其基因多态性的研究. 遗传, 2004, 26(6): 807-810.
[5] Bazzoni F, Beutler B. The tumor necrosis factor ligand and receptor families. N Engl J Med, 1996, 334(26): 1717-1725.
[6] García-Castillo J, Pelegrín P, Mulero V, Meseguer J. Molecular cloning and expression analysis of tumor necrosis factor a from a marine fish reveal its constitutive expression and ubiquitous nature. Immunogenetics, 2002, 54(3): 200-207.
[7] Pfeffer K. Biological functions of tumor necrosis factor cytokines and their receptors. Cytokine Growth Factor Rev, 2003, 14(3-4): 185-191.
[8] Hehlgans T, Pfeffer K. The intriguing biology of the tumour necrosis factor/tumour necrosis factor receptor superfamily: players, rules and the games. Immunology, 2005, 115(1): 1-20.
[9] Tang XR, Fenton MJ, Amar S. Identification and functional characterization of a novel binding site on TNF-α promoter. Proc Natl Acad Sci USA, 2003, 100(7): 4096-4101.
[10] Hong YH, Lillehoj HS, Lee SH, Park DW, Lillehoj EP. Molecular cloning and characterization of chicken lipopolysaccharide-induced TNF-a factor (LITAF). Dev Comp Immunol, 2006, 30(10): 919-929.
[11] Yu YD, Qiu LM, Song LS, Zhao JM, Ni DJ, Zhang Y. Molecular cloning and characterization of a putative lipopolysaccharide-induced TNF-alpha factor (LITAF) gene homologue from Zhikong scallop Chlamys farreri. Fish Shellfish Immunol, 2006, 23(2): 419-429.
[12] Park EM, Kim YO, Nam BH, Kong HJ, Kim WJ, Lee SJ, Kong IS, Choi TJ. Cloning, characterization and expression analysis of the gene for a putative lipopolysaccha-ride-induced TNF-alpha factor of the Pacific oyster, Crassostrea gigas. Fish Shellfish Immunol, 2008, 24(1): 11-17.
[13] Zhang DC, Jiang JJ, Jiang SG, Ma JJ, Su TF, Qiu LH, Zhu CY, Xu HP. Molecular characterization and expression analysis of a putative LPS-induced TNF-alpha factor (LITAF) from pearl oyster Pinctada fucata. Fish Shellfish Immunol, 2009, 27(3): 391-396.
[14] Tang XR, Marciano DL, Leeman SE, Amar S. LPS induces the interaction of a transcription factor, LPS-induced TNF alpha factor, and STAT6(B) with effects on multiple cy-tokines. Proc Natl Acad Sci USA, 2005, 102(14): 5132- 5137.
[15] Nagashima K, Sato M, Kawamata K, Nakamura A, Ohta T. Genetic structure of Japanese scallop population in Hok--kaido, analyzed by mitochondrial haplotype distribution. Mar Biotechno, 2005, 7(1): 1-10.
[16] 刘卫东, 鲍相渤, 宋文涛, 周遵春, 赫崇波, 虞星炬. 虾夷扇贝遗传连锁图谱的初步构建. 遗传, 2009, 31(6): 629-637.
[17] Ponting CP, Mott R, Bork P, Copley RR. Novel protein domains and repeats in Drosophila melanogaster: insights into structure, function, and evolution. Ge-nome Res, 2001, 11(12): 1996-2008.
[18] Bolcato-Bellemin AL, Mattei MG, Fenton M, Amar S. Molecular cloning and characterization of mouse LITAF cDNA: role in the regulation of tumor necrosis factor-alpha (TNF-alpha) gene expression. J Endotoxin Res, 2004, 10(1): 15-23.
[19] Li HJ, Yang Q, Gao XG, Su H, Wang J, He CB. Identification and expression of a putative LPS-induced TNF-α factor from Asiatic hard clam Meretrix meretrix. Mol Biol Rep, 2012, 39(2): 865-871.
[20] Culliney JL. Larval development of the giant scallop Placopecten magellanicus (Gmelin). Biol Bull, 1974, 147(2): 321-332.
[21] Lacoste A, De Cian MC, Cu