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

免疫调节因子TIPE2慢病毒载体的构建及病毒包装

Construction of human immune regulator TIPE2 lentiviral vector and lentivirus packaging
下载PDF
导出
摘要 目的:TIPE2[Tumor necrosis factor(TNF)-αinduced protein 8-like 2(TNFAIP8L2),TIPE2]慢病毒载体的构建及病毒包装。方法:首先构建带有3×FLAG标签的pCDNA3.1-3×FLAG-TIPE2质粒,确认表达后再将标签连同TIPE2序列一起克隆入pCDH-CMV-MCS-EF1-Puro慢病毒表达载体,然后与pMD2.G、psPAX2共转染293T细胞并收获上清,进一步用含病毒颗粒的上清感染PLC/PRF/5及HEK293T细胞,Western blot检测感染后TIPE2蛋白表达情况。结果:pCDNA3.1-3×FLAGTIPE2及pCDH-CMV-MCS-EF1-Puro-TIPE2质粒构建成功,并能最终产出具有感染性的病毒颗粒。结论:成功构建了TIPE2慢病毒载体并获得具有感染性的病毒颗粒,为进一步研究TIPE2功能提供了有力的支持。 Objective：To package lentivirus carrying human TIPE2 （Tumor necrosis factor （TNF）-ct induced protein 8-like 2 （TNFAIP8L2）, TIPE2） gene. Methods： The human TIPE2 gene was cloned into pDNA3. 1-3 x FLAG vector to generate pCDNA3.1- 3× FLAG-TIPE2 plasmid, after sequencing, 3× FLAG-TIPE2 fragment was inserted into pCDH-CMV-MCS-EF1-Puro lentiviral vector. The pCDH-CMV-MCS-EFI-Puro/3× FLAG-TIPE2 plasmids were co-transfected with pMD2. G,psPAX2 plasmids into HEK293T cells to harvest infectious virus particles, followed by infecting PLC/PRF/5 and HEK293T cells. Results： The TIPE2 expressing lentiviral vector was constructed successfully, and finally got infectious virus particles. Conclusion： The human TIPE2 recombinant lentiviral vector was successfully constructed and packaged.
作者 王耀辉 娄强 陈九格 张军 马远方
机构地区 河南大学医学院抗体药物河南省工程实验室
出处 《中国免疫学杂志》 CAS CSCD 北大核心 2013年第10期1077-1079,共3页 Chinese Journal of Immunology
基金 国家自然科学基金资助(30972687)
关键词 免疫调节因子 TIPE2 慢病毒 病毒包装 Immune regulator TIPE2 Lentivirus Virus package
分类号 R392-3 [医药卫生—免疫学]
  • 相关文献

参考文献12

  • 1Sun H, Gong S, Carmody R J et al. TIPE2, a negative regulator of innate and adaptive immunity that maintains immune homeostasis [J] . Cell, 2008; 133 (3) :415426.
  • 2Zhang G, Hao C, Xi W et al. Tissue-specific expression of TIPE2 provides insights into its function [J]. Mol Immunol, 2010; 47 ( 15) : 2435-2442.
  • 3Brautbar Y G, Johnson D, Zhang L et al. The anti-inflammatory TIPE2 is an inhibitor of the oncogenic Ras [J]. Mol Cell, 2012; 45 (5): 610-618.
  • 4Zhang L, Shia Y, Wang Y et al. The unique expression profile of hu?man TIPE2 suggests new functions beyond its role in immune regula?tionj J]. Mol Immunol, 2011 ; 48 (9-10) : 1209-1215.
  • 5Li D, Song L, Fan Y et al. Down-regulation ofTIPE2 mRNA expres?sion in peripheral blood mononuclear cells from patients with systemic lupus erythematosus[J]. Clin Immunol, 2009; 133(3): 422427.
  • 6Zhang S, Zhang Y, Wei X et al. Expression and regulation of a novel identified TNF AlPS family is associated with diabetic nephropathy [J]. Biochim Biophys Acta, 2010; 1802(11) :1078-1086.
  • 7Xi W, Hu Y, Liu Y et al. Roles of TIPE2 in hepatitis B virusinduced hepatic inflammation in humans and mice [J]. Mol Immunol, 2011 ; 48(9-10): 1203-1208.
  • 8Zhang Y, Wei X, Liu L et al. TIPE2, a novel regulator of immunity, protects against experimental stroke[J]. J Bioi Chern, 2012; 287 (39) :32546-32555.
  • 9毛颖佳,郑源强,石艳春.慢病毒载体及其应用的研究进展[J].中国生物制品学杂志,2009,22(2):196-200. 被引量：13
  • 10Kock N, Kasmieh R, Weissleder R et al. Tumor therapy mediated by lentiviral expression of shBcl-2 and S- TRAIL [J]. Neoplasia, 2007; 9(5) :435442.

二级参考文献37

  • 1石艳春,梁浩,旭日干.融合表达IL-2和EGFP逆转录病毒载体的构建[J].中国生物制品学杂志,2006,19(1):1-4. 被引量：3
  • 2Aris K, Michael T, Oliver G, et al. Discovery and analysis of the first endogenous lentivirus. PNAS, 2007, 104 (15): 6261-6265.
  • 3Parolin C, Sodroski L A defective HIV-1 vector for gene transfer to human lymphocytes. J Mol Med, 1995, 73 (6): 279-288.
  • 4Curran MA, Kaiser SM, Achacoso PL, et al. Efficient transduction of nondividing cells by optimized feline immunodeficiency virus vectors. Mol Ther, 2000, 1 ( 1 ): 31-38.
  • 5Mitrophanous K, Yoon S, Rohll J, etal. Stable gene transfer to the nervous system using a non-primate lentiviral vector. Gene Ther. 1999, 6 (11): 1808-1818.
  • 6Mselli-Lakhal L, Favier C, Da Silva Teixeira MF, et al. Defective RNA packaging is responsible for low transduction efficiency of CAEV-based vectors. Arch Virol, 1998, 143 (4): 681-695.
  • 7Berkowitz R, Ilves H, Lin WY, etal. Construction and molecular analysis of gene transfer systems derived from bovine immunodeficiency virus. J Virol, 2001, 75 (7): 3371-3382.
  • 8Berkowitz RD, Ilves H, Plavec I, et al. Gene transfer systems derived from Visna virus: analysis of virus production and infectivity. Virology, 2001, 279 (1): 116-129.
  • 9Poeschla EM. Non-primate lentiviral vectors. Curr Opin Mol Ther, 2003, 5 (5): 529-540.
  • 10Cockrell AS, Kafri T. Gene delivery by lentivirus vectors. Mol Biotechnol, 2007, 36(3): 184-204.

共引文献12

中国免疫学杂志
2013年 第10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