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

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

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

赫崇波^1,2, 朱宝¹, 刘卫东², 鲍相渤², 李云峰², 单忠国³, 李宏俊⁴

1. 辽宁师范大学生命科学学院, 大连 116029 2. 辽宁省海洋水产科学研究院, 辽宁省海洋水产分子生物学重点实验室, 大连 116023 3. 厦门大学海洋与环境学院, 厦门 361005 4. 国家海洋环境监测中心, 大连 116023

收稿日期:2011-10-28 修回日期:2012-02-21 出版日期:2012-06-20 发布日期:2012-06-25
通讯作者: 赫崇波 E-mail:hechongbo@hotmail.com
基金资助:
国家海洋公益性行业科研专项(编号：200805037)和国家自然科学基金项目(编号：31140073)资助

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

摘要/Abstract

摘要： 脂多糖诱导的肿瘤坏死因子(Lipopolysaccharide-induced TNF-alpha factor, LITAF)是一类重要的炎症细胞因子, 在先天性免疫系统中发挥重要的介质作用。文章根据虾夷扇贝LITAF基因EST序列, 应用RACE技术克隆了虾夷扇贝LITAF全长cDNA, 对序列及编码的氨基酸进行生物信息学分析。结果显示, 该基因cDNA全长1 551 bp, 其5′非编码区包含76 bp, 3′非编码区包含1 001 bp; 开放阅读框(ORF)为474 bp, 编码157个氨基酸, 氨基酸序列中存在一个保守的LITAF结构域; 理论分子量16.99 kDa, 等电点为6.24。LITAF基因序列为3 698 bp, 由3个外显子和两个内含子组成。利用实时荧光定量PCR技术分析LITAF在虾夷扇贝不同组织、不同胚胎发育阶段以及鳗弧菌(Vibrio anguillarum)刺激后各时间段的表达情况。结果表明：LITAF基因在所检测的6个成体组织中均有表达, 其中肾脏的表达量最高; 胚胎发育的7个时期中, 担轮幼体时期表达量最高; 菌刺激36 h实验组与对照组的表达量差异大。LITAF基因是LITAF家族的一员, 推测LITAF基因参与虾夷扇贝的先天性免疫反应。

关键词: 虾夷扇贝, 脂多糖诱导的肿瘤坏死因子, 序列分析, 胚胎发育, 基因表达

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

赫崇波，朱宝，刘卫东，鲍相渤，李云峰，单忠国，李宏俊. 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[J]. 遗传, 2012, 34(6): 736-741.

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.

参考文献

[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

编辑推荐

Metrics

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

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

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

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	王舜泽, 江丰, 朱东丽, 杨铁林, 郭燕. Hi-C技术在三维基因组学和疾病致病机理研究中的应用[J]. 遗传, 2023, 45(4): 279-294.
[2]	王文龙, 张春霞. 哺乳动物卵子与早期胚胎中全转录组poly(A)尾研究进展[J]. 遗传, 2023, 45(4): 273-278.
[3]	徐思远, 寿佳, 吴强. HS5-1增强子eRNA PEARL对原钙粘蛋白α基因簇的表达调控[J]. 遗传, 2022, 44(8): 695-764.
[4]	赵岩, 王晨鑫, 杨天明, 李春爽, 张丽宏, 杜冬妮, 王若曦, 王静, 魏民, 巴雪青. DNA氧化损伤8-羟鸟嘌呤与肿瘤的发生发展[J]. 遗传, 2022, 44(6): 466-477.
[5]	田子钊, 周晨希, 周伟, 李沫, 褚云鹏, 怀聪, 秦胜营. 通过时间基因表达谱分析探究异烟肼引起肝损伤的机制[J]. 遗传, 2022, 44(6): 501-509.
[6]	王梓川, 张嘉祺, 李磊. 哺乳动物早期胚胎发育的体外研究[J]. 遗传, 2022, 44(4): 269-274.
[7]	朱前彬, 甘志承, 李晓翠, 张英杰, 赵合明, 黄先忠. 小鼠耳芥MAPKKK基因家族全基因组鉴定及进化与表达[J]. 遗传, 2022, 44(11): 1044-1055.
[8]	周聪, 周强伟, 成盛, 李国亮. CTCF在介导三维基因组形成及调控基因表达中的研究进展[J]. 遗传, 2021, 43(9): 816-821.
[9]	胡广伟, 张珍珍, 高焕. 文昌鱼左右体轴建立机制的研究进展[J]. 遗传, 2021, 43(2): 134-141.
[10]	徐海冬, 宁博林, 牟芳, 李辉, 王宁. 选择性多聚腺苷酸化的生物学效应及其调控机制研究进展[J]. 遗传, 2021, 43(1): 4-15.
[11]	王涛涛, 杨勇, 魏唯, 林辰涛, 马留银. 互花米草NAC转录因子家族的鉴定与表达分析[J]. 遗传, 2020, 42(2): 194-211.
[12]	陈会友, 张建敏, 李柏森, 邓永琳, 张龚炜. 犏牛雄性不育的减数分裂基因表达与表观遗传调控研究进展[J]. 遗传, 2020, 42(11): 1081-1092.
[13]	高晓萌, 张治华. 生物大分子“液-液相分离”调控染色质三维空间结构和功能[J]. 遗传, 2020, 42(1): 45-56.
[14]	禹奇超,宋彬,邹轩轩,王岭,刘德权,李波,马昆. 乳腺癌癌旁组织特异性表达基因分析[J]. 遗传, 2019, 41(7): 625-633.
[15]	石田培,张莉. 全转录组学在畜牧业中的应用[J]. 遗传, 2019, 41(3): 193-205.