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

hIL-18基因真核表达载体的构建及序列分析

The Construction of Eukaryotic Expression Vecto of hIL-18 Gene and Its Sequence Analysis
下载PDF
导出
摘要 在获得人白细胞介素18(hIL-18)基因重组质粒pGEM-T-hIL-18的基础上,用EcoRⅠ、SalⅠ双酶切该质粒,回收小片段hIL-18基因;同时双酶切真核表达质粒pECFP-C1,回收大片段,二者连接转化大肠埃希菌感受态细胞。对重组质粒pECFP-C1-hIL-18进行PCR扩增、酶切鉴定及序列测定,测序结果表明,克隆至pGEM-T中的IL-18 cDNA包含成熟IL-18的全部编码序列。本研究获得了融合有报告基因pECFP-C1的真核表达载体,为下一步进行hIL-18在真核细胞中的表达打下了良好的基础,亦为在基因水平上探讨hIL-18的生物学功能提供了物质基础。 Recombinant plasmid pGEM-T-hIL-18 was constructed, then the plasmid pGEM-T-hIL-18 and pECFP-C1 was enzymed by EcoR Ⅰ and Sal Ⅰ , then was transformed into the E. coli strain DH5αto construct recombinant plasmid pECFP-CI-hIL-18. The recombinant plasmid was amplified by pcr,enzymed by EcoR I and Sal I and the sequence was analyzed by computer software. It was confirmed that in the sequence of pECFP-CI-hIL-18 contains all the coding sequence for mature protein of hIL-18. We obtained hIL-18 gene and constructed pECFP-CI-hIL-18. The results laid a solid foundation and prepared experimental material for the future of eukaryotic expression and bioactivity of IL-18.
作者 纪丽丽 杨玉英 李士泽
机构地区 山东畜牧兽医职业学院 黑龙江八一农垦大学
出处 《动物医学进展》 CSCD 2007年第9期45-48,共4页 Progress In Veterinary Medicine
基金 黑龙江省教育厅科研项目(10541157)
关键词 白细胞介素18 真核载体 序列测定 Interleukin-18 eukaryotic expression vector sequence analysis
分类号 Q78 [生物学—分子生物学]
  • 相关文献

参考文献8

  • 1Ushio S,Namba M,Okura T.et al.Cloning of the cDNA for human IFN-gamma-inducting factor expression in Escherichia coil and studies On the biologic activities of the protein[J].J Immunol,1996,156:4274-4279.
  • 2纪丽丽,杨玉英,李士泽,闫邦样.重组人白细胞介素18基因的克隆及序列测定[J].黑龙江八一农垦大学学报,2005,17(5):61-64. 被引量：1
  • 3Morgan R A,Anderson N.Human gene therapy[J].Annu Rev Biochem,1993,62:191-204.
  • 4Yang T T,Cheng L,Kain S R.Optimized codon usage and chromophore mutations provide enhanced sensitivity with the green fluorescent protein[J].Nucleic Acids Rea,1996,24(22):4592-4593.
  • 5Takeda K,Tsutsui H,Ynshimoto T,et al.Defective NK cell activity and Th1 response in IL-18-deficient mice[J].Immunity,1998,(8):383-390.
  • 6Wang Q,Yu H,Zhang L,et al.Vaccination with IL-18 genemodified superantigen-coatedtumor cells elicits potent antitumor immune respons[J].J Cancer Res Clin Oncol,2001,127(12):718-726.
  • 7Hegardt P,Widegren B,Li L,et al.Nitric oxide synthase inhibitor and IL-18 enhance the anti-tumor immune response of rats carrying an intrahepatic colon carcinoma[J].Cancer Immunol Immunother,2001,50(9):491-501.
  • 8纪丽丽,王玉艳,王纯净,李士泽,杨玉英.白细胞介素-18及其生物学作用[J].黑龙江畜牧兽医,2005(12):76-78. 被引量：4

二级参考文献25

  • 1Okamura H, Nagata K, Komatsu T, et al. A novel costimulatory factor for gamma interferon induction found in the livers of mice causes endotoxic shock[J]. Infect Immune, 1995, 63 (10): 3966-3972.
  • 2Ushio S, Namba M, Okura T, et al. Cloning of the cDNA for human IFN-gamma-inducting factor expression in Escherichia coil and studies on thebiologic activities of theprotein[J].J Immunol, 1996, (156):4274-4279.
  • 3Stoll S, Jonuleit H, Schmitt E, et al. Production of functional IL-18 by different subtypes of murine and human dendriticcells(DC): DC derived IL-18 enhances IL-12 independent Thl development[J].Eur J Immunol, 1998, 28 (10): 3231-3239.
  • 4Tanaka F, Hshimoto W, Okamura H, et al. Rapid generation of potent tumor specific cytotoxic T lymphocytes by interleukin 18 using dendritic cells and natural killer cells[J]. Cancer Res, 2000, 60 (17):4838-4844.
  • 5Osaki T, Peron J M, Cai Q, et al. IFN-γ inducing factor 18 and administration mediates IFN-γ and IL-12 in dependent antitumor effects[J].J Immunol, 1998, 160 (4): 1742-1749.
  • 6Kohno [, [ataoka J, Ohtsuki T et al IFN-gamma-inducing factor(IGIF) is a costimulatory factor on the activation of Thl but not Th2 cells and exerts its effect independently of IL-12[J].J Immunol,1997, (158):1541-1550.
  • 7Dao T, Ohasi K, Kayano T, et al. Interferon-γ-inducing factor, a novel cytokine, enhances Fas ligand-mediated cytotoxicity ofmurine T helper 1 ce11[J]. Celluar Immunology, 1996, (173): 230-235.
  • 8GuY, KuidaK, Tsutsui H, et al. Activation of Interferon-gamma Inducing Factor Mediated by Interleukin-lbeta Converting Enzyme[J].Science, 1997, (275): 206-209.
  • 9Okamura H, Nagata K, Komatsu T,et al. A novel costimulatory factor for gamma interferon induction found in the livers of mice causes endotoxic shock[J]. Infect Immune, 1995 ,63 (10) :3966 -3972.
  • 10Ushio S, Namba M, Okura T,et al. Cloning of the eDNA for human IFN - gamma - inducting factor expression in Escherichia coil and studies on the biologic activities of the protein [J]. J Immunol,1996,156:4274 -4279.

共引文献3

动物医学进展
2007年 第9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