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纤维连结蛋白启动子的克隆及转录活性的研究

Cloning of the fibronectin promoter and observation of its transcriptional activity
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摘要 目的:克隆纤维连结蛋白(FN)启动子和检测其转录活性,并初步研究增加SV40增强子对其转录活性的影响。方法:从正常人gDNA中克隆FN启动子,驱动氯霉素乙酰转移酶报告基因表达,构建成有SV40增强子和无SV40增强子两个重组体报告载体。利用脂质体介导的转染技术导入HT1080细胞,检测各重组体的瞬时表达,确定克隆FN启动子转录活性和SV40增强子对其转录活性的影响。结果:克隆的FN启动子在HT1080细胞中活性比SV40启动子稍强,在加入SV40增强子时活性提高了约6倍。结论:成功的克隆FN启动子,在HT1080细胞中有活性,通过增加SV40增强子能大大提高其活性,为运用FN启动子和特异性靶细胞治疗各种遗传性疾病提供了一定基础。 AIM： To clone fibronectin （FN） promoter and identify its transcriptional activity combined with SV40 enhancer. METHODS： FN promoter was amplified from genomic DNA by PCR and cloned into pCat3 - basic and pCat3 - ehancer vector to drive CAT expression. They were transferred to HT1080 cells with lipofectin. Then the instant CAT expression of different plasids was detected and the FN promoter transcriptional activity and its activity when it was combined with SV40 enhancer were evaluated. RESULTS： The FN promoter was cloned successfully. Its transcriptional activity was slightly stronger than the activity of SV40 promoter. Combined with SV40 enhancer, its activity was improved by about six times. CONCLUSION： The FN promoter cloned by us has transcriptional activity. The activity is increased by adding SV40 enhancer. These will provide useful information for gene therapy for combined FN promoter to cure some heredity diseases.
作者 李奎 唐俊明 王家宁 王明江 黄永章
机构地区 郧阳医学院附属人民医院临床医学研究所 郧阳医学院生理教研室
出处 《中国病理生理杂志》 CAS CSCD 北大核心 2006年第7期1272-1276,共5页 Chinese Journal of Pathophysiology
基金 湖北省自然科学基金资助项目(2005ABA079)
关键词 纤维连结蛋白启动子 克隆 分子 SV40增强子 基因 报告 Fibronectin promoter Cloning, molecular SV40 enhancer Genes, reporter
分类号 R363 [医药卫生—病理学]
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参考文献15

  • 1何祥梁,何东华.肺癌基因治疗研究进展[J].中国病理生理杂志,2002,18(11):1447-1450. 被引量：2
  • 2Ramesh N,Lau S,Palmer TD,etal.High-level human adenosine deaminase expression in dog skin fibroblasts is not sustainedfollowing transplantation[J].Human Gene Ther,1993,4(1):3-7.
  • 3Xu L,Gao C,Sands MS,et al.Neonatal or hepatocyte growth factor-potentiated adult gene therapy with a retroviral vector results in therapeutic levels of canine factor Ⅸ for hemophilia B[J].Blood,2003,101 (10):3924 -3932.
  • 4Moullier P,Bohl D,Cardoso J,et al.Long-term delivery of a lysosomal enzyme by genetically modified fibroblasts in dogs[J].Nature Med,1995,1(4):353 -357.
  • 5Baek SC,Lin Q,Robbins PB,et al.Sustainable systemic delivery via a single injection of lentivirus into human skin tissue[J].Hum Gene Ther,2001,12(12):1551 -1558.
  • 6Palmer TD,Rosman G J,Osborne WR,et al.Genetically midified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes[J].Proc Natl Acad Sci USA,1991,88(4):1330-1334.
  • 7Nakajima T,Nakamura T,Tsunoda S,et al.E1A -responsive elements for repression of rat fibronectin gene transcription[J].Mol Cell Biol,1992,12(6):2837 -2846.
  • 8Taylor RM,Wolfe JH.Decreased lysosomal storage in the aldult MPS Ⅶ mpuse brain in the vicinity of graft of retroviral vector-corrected fibroblasts secreting gigh levels of β-glucuronidase[J].Nature Med,1997,3(7):771-774.
  • 9Dean DC,Bowlus CL,Bourgeois S,et al.Cloning and analysis of the promoter region of the human fibronectin gene[J].Proc Natl Acad Sci USA,1987,84(7):1876.-1880.
  • 10段山,方小武,陈路明,曾瑞萍,杜传书.人β地中海贫血IVSⅡ654(C→T)突变基因的克隆和真核表达体系的构建[J].中国病理生理杂志,2003,19(1):22-26. 被引量：1

二级参考文献20

  • 1茅矛,杜传书,王菁,陈路明.地中海贫血的红细胞抗氧化酶和核苷酸代谢酶活性变化[J].中国病理生理杂志,1994,10(5):477-481. 被引量：8
  • 2[1]Takayama K, Nakanishi Y, Hara N,et al. Gene therapy for lung cancer treatment [J]. Nippon Rinsho,2000,58(5):1048-1052.
  • 3[2]Pearson AS, Spitz FR, Swisher SG,et al. Up-regulation of the proapoptotic mediators Bax and Bak after adenovirus-mediated TP53 gene transfer in lung cancer cells[J]. Clin Cancer Res,2000,6(3):887-890.
  • 4[3]Nemunaitis J,Swisher SG, Timmons T,et al. Adenovirus-mediated TP53 gene transfer in sequence with cisplatin to tumors of patients with non-small-cell lung cancer[J]. J Clin Oncol,2000,18(3):609-622.
  • 5[4]Gai M. gene in human lung cancer cells after retroviral-mediated gene transfer[J]. Hum Gene Ther,1993,4(5):617-622.
  • 6[5]Inoue A, Narumi K, Matsubara N,et al. Administration of wild-type TP53 adenoviral vector synergistically enhances the cytotoxicity of anti-cancer drugs in human lung cancer cells irrespective of the status of TP53 gene[J]. Cancer Lett,2000,157(1):105-112.
  • 7[6]Yamanishi Y, Hiyama K,Maeda H,et al. Specific growth inhibition of SCLC cells by adenovirus vector expressing antisense c-kit transcripts[J].Jpn J Cancer Res,1996,87(5):534-540.
  • 8[7]Lee GT, Wu S, Gabrllovich D,et al. Antitumor effects of adenovirus expressing antisense insulin-like growth factor I on human lung cancer cell lines[J].Cancer Res,1996,56(7):3038-3043.
  • 9[8]Moolten FL. Tumor chemosensitivity conferred by inserted herpes thymidene kinase genes; Paradigm for a prospective cancer control strategy[J]. Cancer Res,1986,46(10):5276-5281.
  • 10[9]Nishino K, Osaki T, Kumagai T,et al.Adenovirus-mediated gene therapy specific for small cell lung cancer cells using a Myc-Max binding motif[J]. Int J Cancer,2001,91(6):851-856.

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中国病理生理杂志
2006年 第7期

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