期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

人天然免疫基因BCL10的猪同源物的识别、克隆与初步表达分析 被引量：5

In silico identification,molecular cloning and verification of a novel pig gene homologous to human BCL10 of in-nate immunity and its preliminary expression profiles in pigs
下载PDF
导出
摘要 采用电子克隆方法克隆到大小为925bp的人天然免疫蛋白BCL10的猪同源基因完整cDNA序列(GenBank登录号:EU088132),并利用RT-PCR方法从猪的全血中扩增出包含702bp的完整开放读码框架(ORF)的cDNA片段。经核酸测序,证明与电子克隆结果相符。利用NCBIBLAST分析该cDNA包含3个大小为57bp、289bp和356bp的外显子,并且定位于猪的4号染色体上。采用半定量PCR技术检测基础水平猪各组织BCL10基因mRNA表达丰度,并将该基因构建到带有绿色标签的真核表达载体pEGFP-C1中,采用脂质体转染法将该基因转入PK-15细胞,通过绿色荧光标记和RT-PCR方法检测实验组的BCL10蛋白表达。研究结果表明,BCL10基因mRNA在脾脏中表达最高;胸腺、大脑和淋巴结表达次之,而肝脏只有微量表达,肾脏没有检测到表达;同时BCL10基因在PK-15细胞中得到了有效表达。 We identified and characterized a novel swine gene Bcl10 with GenBank （Accession No. EU088132） which was homologous to human BCLIO （B-cell CLL/lymphoma 10） gene. The full-length cDNA of 925 bp for swine BCLIO was in silico cloned, and then its ORF of 702 bp coding 233 amino acid residues was verified by RT-PCR and DNA sequencing. NCBI BLAST assay indicated that this cDNA contained three extrons with a length of 57, 289 and 356 bp respectively, and it was located on the chromosome 4 of pig genome. Using semi-quantitative PCR, preliminary expression profiles of swineBCLIO were verified in different swine tissues. The expression of swine BCLIO was verified by GFP marker and RT-PCR assay. We found that, BCLIO expressed in high level in swine spleen, and with a modest level in thymus, brain and lymph node; low level mRNA was expressed in liver and not detectable level in kidney. The swine BCLIO gene was cloned to the GFP-containing eukaryotic expression vector pEGFP-C1 and transfected to PK-15 cell line by lipofectin. BCLIO was expressed effectively in PK-15 cells. In summary, we cloned a novel swine BCLIO gene, and investigated its expression characters. This will be the fundament of the future research on its function.
作者 何建锋 张彦明 段会娟 曹金锁 张德礼
机构地区 西北农林科技大学动物医学院预防兽医学系病毒免疫与生物信息研究室
出处 《遗传》 CAS CSCD 北大核心 2008年第6期747-754,共8页 Hereditas（Beijing)
基金 西北农林科技大学引进人才基金资助项目(编号:01140406) 国家自然科学基金项目(编号:30270342)资助~~
关键词 BCL10基因 克隆 表达分析 真核表达 BCL10 gene cloning expression analysis eukaryotic expression
分类号 Q78 [生物学—分子生物学]
  • 相关文献

参考文献19

  • 1Zhang Q, Siebert R, Yan M, Hinzmann B, Cui X, Xue L, Rakestraw KM, Naeve CW, Beckmann G, Weisenburger DD, Sanger WG, Nowotny H, Vesely M, Callet-Bauchu E,Salles G, Dixit VM, Rosenthal A, Schlegelberger B, Morris SW. Inactivating mutations and overexpression of BCL10, a caspase recruitment domain-containing gene in MALT lymphoma with t(1;14)(p22;q32). Nat Genet, 1999, 22: 63-68.
  • 2Maes B, Demunter A, Peeters B, De Wolf-Peeters C. BCL10 mutation does not represent an important pathogenic mechanism in gastric MALT-type lymphoma, and- the presence of the API2-MLT fusion is associated withaberrant nuclear BCL10 expression. Blood, 2002, 99: 1398-1404.
  • 3Sagaert X, Laurent M, Baens M, Wlodarska I, De Wolf-Peeters C. MALT1 and BCL10 aberrations in MALT lymphomas and their effect on the expression of BCL10 in the tumour cells. Mod Pathol, 2006, 19(2): 225-232.
  • 4Bose S, Goldman JM, Melo JV. Mutations of the BCL10 gene are not associated with the blast crisis of chronic myeloid leukemia. Leukemia, 1999, 13: 1894-1896.
  • 5Lucas PC, Yonezumi M, Inohara N, McAllister-Lucas LM, Abazeed ME, Chen FF, Yamaoka S, Seto M, Nunez G. BCL10 and MALT1, independent targets of chromosomal translocation in MALT lymphoma, cooperate in a novel NF-kB signaling pathway. J Biol Chem, 2001, 276(22): 19012-19019.
  • 6Willis TG, Jadayel DM, Du MQ, Peng H, Perry AR, Abdul-Rauf M, Price H, Karran L, Majekodunmi O, Wlodarska I, Pan L, Crook T, Hamoudi R, Isaacson PG, Dyer MJ. BCL10 is involved in t (1:14)(p22; q32) of MALT B cell lymphoma and mutated inmultiple tumor types. Cell, 1999, 96: 35-45.
  • 7Thome M. CARMA1, BCL-10 and MALT1 in lymphocyte development and activation. Nat Rev Immunol, 2004, 4: 348-359.
  • 8张德礼,丁培国,凌伦奖,陈润生,马大龙.人类新基因C17orf32的电子克隆和编码区序列RT-PCR验证[J].生物化学与生物物理进展,2002,29(4):543-549. 被引量：22
  • 9张华莉,邓昊,张瑞芳,夏昆,夏家辉.人类TECTB基因的电子克隆[J].Acta Genetica Sinica,2003,30(4):317-320. 被引量：17
  • 10张德礼,孙晓静,凌伦奖,陈润生,马大龙.人类SR蛋白超家族新成员——SFRS12(SRrp508)的基因克隆和特性分析[J].Acta Genetica Sinica,2002,29(5):377-383. 被引量：43

二级参考文献46

  • 1Van Rijin P A. A preliminary map ofepitopes on envelope glycoprotein E1 of HCV strain Brescia[J].Veterinary Microbiology, 1992,33:212～230.
  • 2Van Rijin P A. Antigen structure of envelope glycoprotein E1 of hog cholera virus[J].Journal of general virology, 1994,68:3934～3942.
  • 3Van Rijn P A, Bossers A, Wensvoort G, Moormann R J M. Classicalswine fever virus envelope glycoprotein in E2 containing one structural antigenic unit protects swine from lethal CSFV challenge [J].J Gen Virol,1996,77:2737～2745.
  • 4Hammond J M, Jansen E S, Morrissy C J. Protection of pigs against in contact challenge with classical swine fever following oral or subcutaneous vaccination with a recombinant porcine adenovirus.Virus[J].Res,2003, 97(2):151～157.
  • 5J萨姆布鲁克 E.F.弗里奇 T.曼尼阿蒂斯.分子克隆实验指南[M](第2版)[M].北京:科学出版社,1995.10-18.
  • 6Zhang S, Zubay G, Goldman E. Low-usage codons in Escherichia coli,yeast, fruit fly and primates[J].Gene, 1991,105:61～72.
  • 7Sharp P M, Li W H. Codon Usage in Regulatory Genes in Escherichia Coli Does Not Reflect Selection for rare Codons [J].Nucleic Acids Res,1986,14( 19): 7737～7749.
  • 8Ikemura T. Correlation Between the Abundance of Escherichia Coli Transfer RNAs and the Occurrence of the Respective Codons in Its Protein Genes[J].J Mol Biol,1981,146(1):1～21.
  • 9Meyers G, Thiel H J. Molecular characterization of pestivirus[J]. Adv Virus Res, 1996, 47:53～118.
  • 10Beate M, Kummerer, Myers G. Correlation between Point Mutations in NS2 and the Viability and Cytopathogenicity of Bovine Viral Diarrhea Virus Strain Oregon Analyzed with an Infectious cDNA Clone [J].Virol, 2000, 74:390～400.

共引文献157

同被引文献26

  • 1Mao X, Hamoudi R A. Molecular and cytogenetic anal- ysis of gliohlastoma multiforme[J]. Cancer Genet Cy- togenet, 2000,122 ( 2 ) : 87-92.
  • 2Willis T G,Jadayel D M,Peng H,et al. Bcl10 is in- volved in t(1;14)(p22 ;q32) of MALT B cell lympho- ma and mutated in multiple tumor types[J]. Cell, 1999,96(1):35-45.
  • 3Wang L,Guo Y, Huang W J,et al. Card10 is a novel caspase recruitment domain/membrane-associated guanylate kinase family .member that interacts with BCL10 and activates NF-kappa B[J]. J Biol Chem, 2001,276(24) :21405-21409.
  • 4Guo B,Su T T,Rawlings D J. Protein kinase C family functions in B-cell activation[J]. Curr Opin Immunol,2004,16(3):367-373.
  • 5Sun L, Deng L, Ea C K, et al. The TRAF6 ubiquitin ligase and TAK1 kinase mediate IKK activation by BCL10 and MALT1 in T lymphocytes[J]. Mol Cell, 2004,14(3) : 289-301.
  • 6Lin X, Wang D. The roles of CARMA1, Bcl10, and MALT1 in antigen receptor signaling[J]. Semin Im- munol,2004,16(6) :429-435.
  • 7Ishiguro K, Ando T, Goto H, et al. Bcl10 is phospho- rylated on Ser138 by Ca^2+/calmodulin-dependent pro- tein kinase Ⅱ[J]. Mol Immunot, 2007,44 (8) : 2095- 2100.
  • 8Gaide O, Martinon F, Micheau O, et al. Carmal, a CARD-containing binding partner of Bcl10, induces Bcl10 phosphorylation and NF-kappa B activation [J]. Febs Lett,2001,496(2/3) :121-127.
  • 9Sommer K,Guo B,Pomerantz J L,et al. Phosphoryla- tion of the CARMA1 linker controls NF-[kappa] B activation[J]. Immunity,2005,23(6) :561-574.
  • 10Ruland J ,Duncan G S,Elia A,et al. Bcl10 is a positive regulator of antigen receptor-induced activation of NF-kappa B and neural tube elosure[J]. Cell,2001, 104(1) :33-42.

引证文献5

二级引证文献1

遗传
2008年 第6期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部