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

人生存素T34A重组腺病毒载体的构建、鉴定和体外表达 被引量：1

Construction,Identification and Expression of Recombinant Adenovirus Vector for hSurvivin T34A
原文传递
导出
摘要 目的:采用高效简便的GatewayTM系统构建人生存素T34A基因重组腺病毒载体(Ad-hSurvivinT34A)。方法:从pORF9-hSurvivinT34A质粒中克隆人生存素T34A基因,经NcoⅠ/XhoⅠ双酶切后插入到入门载体pEN-TR11的attL1和attL2之间,形成入门克隆;在高效重组酶的作用下,入门克隆与带attR1和attR2位点的病毒骨架载体Ad/CMV/V5-DEST发生体外同源重组,形成表达克隆Ad-hSurvivinT34A。鉴定正确后将其线性化,由Lipo-fectamine2000介导转染293细胞进行病毒包装,收集病毒上清。采用western Blot检测病毒感染的人宫颈癌细胞株Hela细胞中人生存素T34A蛋白的表达。结果:成功构建了人生存素T34A基因的重组腺病毒,该病毒能在细胞中高效表达。结论:本实验利用GatewayTM系统成功构建了Ad-hSurvivinT34A,与传统的腺病毒构建方法相比,更简便和高效,为下一步利用人生存素T34A进行肿瘤免疫基因治疗奠定实验基础。 Objective:To construct a recombinant adenovirus vector with human survivinT34A(Ad-hSurvivinT34A)using GatewayTM system.Methods:The CDS sequence of hSurvivinT34A gene was prepared from plasmid pORF9-hSurvivinT34A,and transferred into pENTR11 within attL1 and attL2 site after double digestion with Nco Ⅰ/Xho Ⅰ to generate the entry clone.Then the entry clone and the target vector Ad/CMV/V5-DEST within attL1 and attL2 sites was recombined together in vivo to create the expression clone Ad-hSurvivinT34A under th...
作者 张鹏 李涛 郎锦义
机构地区 四川省肿瘤医院放射肿瘤中心
出处 《华西医学》 CAS 2008年第6期1362-1364,共3页 West China Medical Journal
关键词 人生存素T34A GatewayTM 重组腺病毒 hSurvivinT34A GatewayTM recombinant adenovirus
分类号 Q78 [生物学—分子生物学]
  • 相关文献

参考文献17

  • 1[1]Altieri DC.Validating survivin as a cancer therapeutic target[J].Nat Rev Cancer,2003,3(1):46-54.
  • 2[2]Adida C,Haioun C,Gaulard P,et al.Prognostic significance of survivin expression in diffuse large B-cell lymphomas[J].Blood,2000,96(5):1921-1925.
  • 3[3]Grossman D,McNiff JM,Li F,et al.Expression and targeting of the apoptosis inhibitor,survivin,in human melanoma[J].J Invest Dermatol,1999,113(6):1076-1081.
  • 4[4]Sloan AE,Parajuli P.Human autologous dendritic cell-glioma fusions:feasibility and capacity to stimulate T cells with proliferative and cytolytic activity[J].J Neurooncol,2003,64(1-2):177-183.
  • 5[5]Yamanaka R,Zullo SA,Ramsey J,et al.Marked enhancement of antitumor immune responses in mouse brain tumor models by genetically modified dendritic cells producing Semliki Forest virus-mediated interleukin-12[J].J Neurosurg,2002,97(3):611-618.
  • 6[6]Yuan ZP,Chen LJ,Fan LY,et al.Liposomal quercetin efficiently suppresses growth of solid tumors in murine models[J].Clin Cancer Res,2006,12(10):3193-3199.
  • 7[7]Andersen MH,Pedersen LO,Becker JC,et al.Identification of a cytotoxic T lymphocyte response to the apoptosis inhibitor protein survivin in cancer patients[J].Cancer Res,2001,61(3):869-872.
  • 8[8]Casati C,Dalerba P,Rivoltini L,et al.The apoptosis inhibitor protein survivin induces tumor-specific CD8+ and CD4+ T cells in colorectal cancer patients[J].Cancer Res,2003,63(15):4507-4515.
  • 9[9]Pisarev V,Yu B,Salup R,et al.Full-length dominant-negative survivin for cancer immunotherapy[J].Clin Cancer Res,2003,9(17):6523-6533;Comment in:6310-6315.
  • 10[10]Altieri DC.Survivin,cancer networks and pathway-directed drug discovery[J].Nat Rev Cancer,2008,8(1):61-70.

同被引文献13

  • 1Lombardi JV,Naji M,Larson RA,et al.Adenoviral mediated ut-eroglobin gene transfer to the adventitia reduces arterial intimal hyperplasia[J].J Surg Res,2001,99:377-380.
  • 2Partieia MK,Natarajan R,Dooley AN,et al.Adenoviral delivery of a leukocyte-type 12 lipoxygenase ribozyme inhibits effects of glucose and platelet-derived growth factor in vascular endothelial and smooth muscle cells[J].Circ Res,2001,88:659-665.
  • 3Rubin A,Mobley B,Hogikyan N,et al.Delivery of an adenoviral vector to the crushed recurrent laryngeal nerve[J].Laryngoseope,2003,113(6):985-989.
  • 4Connelly S,Mech C.Delivery of adenoviral DNA to mouse liver[J].Methods Mol Biol,2004,246:37-52.
  • 5Ma H,Liu Y,Liu S,et al.Oral adeno-associated virus-TRAIL gene therapy suppresses human hepatocellular carcinoma growth in mice[J].Hepatology,2005,42(6):1355-1363.
  • 6Lou J,Manske PR,Aoki M.Adenovirus mediated gene transfer into tendon and tendon sheath[J].J Ort hop Res,1996,14(4):513-517.
  • 7He TC,Zhou S,Costa LT,et al.A simplified system for generation recombinant adenovirus[J].Proc Natl Acad Sci USA,1998,95:2509-2514.
  • 8Ng P,Parks RJ,Cummings DT,et al.An enhanced system for construction of adenoviral vector by the two-plasmid rescue method[J].Hum Gene Ther,2000,11(5):693-699.
  • 9Kolb AF,Siddell SG.Expression of a recombinant monoclonal antibody from a bicistronic mRNA[J].Hybridoma,1997,16:421-426.
  • 10Ja ST,George.Gene therapy progress and prospects:adenoviral vectors[J].Gene Therapy,2003,10:1135-1141.

引证文献1

二级引证文献6

华西医学
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