虾夷扇贝脂多糖诱导的肿瘤坏死因子LITAF基因的克隆及表达分析

doi:10.3724/SP.J.1005.2012.00736

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HEREDITAS ›› 2012, Vol. 34 ›› Issue (6): 736-741.doi: 10.3724/SP.J.1005.2012.00736

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Cloning and expression analysis of lipopolysaccharide-induced TNF-a(LITAF) of Japanese scallop (Mizuhopecten yessoensis)

HE Chong-Bo^1,2, ZHU Bao¹, LIU Wei-Dong², BAO Xiang-Bo², LI Yun-Feng², SHAN Zhong-Guo³, LI Hong-Jun⁴

1. College of Life Science, Liaoning Normal University, Dalian 116029, China 2. Key Lab of Marine Fishery Molecular Biology of Liaoning Province, Liaoning Ocean and Fisheries Science Research Institute, Dalian 116023, China 3. College of Oceanography and Environmental Science, Xiamen University, Dalian 361005, China 4. National Marine Environmental Monitoring Center, Dalian 116023, China

Received:2011-10-28 Revised:2012-02-21 Online:2012-06-20 Published:2012-06-25

Abstract

Abstract: 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.

Key words: Japanese scallop(Mizuhopecten yessoensis), lipopolysaccharide-induced TNF-alpha fac-tor(LITAF), sequence analysis, embryonic development, gene expression

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, 2012, 34(6): 736-741.

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Cloning and expression analysis of lipopolysaccharide-induced TNF-a(LITAF) of Japanese scallop (Mizuhopecten yessoensis)

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