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转录因子CTCF的克隆、表达及其抗体制备 被引量：5

Cloning, expression and antibody preparation of transcription factor CTCF
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摘要 用RT-PCR方法从HeLa细胞总RNA中扩增转录因子CTCF的cDNA序列,克隆至pGEM-TEasy载体上。将CTCF蛋白N段的编码序列亚克隆至pET22b(+)载体中,在大肠杆菌BL21(DE3)中诱导表达,并用镍柱进行亲和纯化。用纯化后的CTCFN段蛋白免疫家兔,获得CTCF的多克隆抗体,进而用于细胞内CTCF蛋白的免疫印迹检测。 CTCF cDNA was amplified from total RNA of HeLa cells and was ligated into pGEM-T Easy plasmid. N domain of CTCF, termed CTCF-N, was expressed in E.coli BL21(DE3) using pET expression system, and then CTCF-N was purified by Ni-NTA agarose column. CTCF-N polyclonal antibody was elicited by immunizing rabbit, then purified and used to detect CTCF level by western blotting.
作者 何莉 朱旭东 毛志勇 朱恒奇 黄培堂
机构地区 军事医学科学院生物工程研究所
出处 《生物技术通讯》 CAS 2004年第5期454-456,共3页 Letters in Biotechnology
关键词 转录因子 蛋白 抗体制备 诱导表达 亚克隆 细胞内 ET 纯化 CDNA序列 总RNA CTCF transcription factor polyclonal antibody western blotting
分类号 Q78 [生物学—分子生物学] R392 [医药卫生—免疫学]
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参考文献8

  • 1Burton T, Liang B, Dibrov A, et al. Transforming growth factor-beta-induced transcription of the Alzheimer beta-amyloid precursor protein gene involves interaction between the CTCF-complex and Smads[J]. Biochem Biophys Res Commun, 2002,295(3):713
  • 2Loukinov DI, Pugacheva E, Vatolin S, et al. BORIS, a novel male germ-line-specific protein associated with epigenetic reprogramming events, shares the same 11-zinc-finger domain with CTCF, the insulator protein involved in reading imprinting marks in the
  • 3Ohlsson R, Renkawitz R, Lobanenkov V. CTCF is a uniquely versatile transcription regulator linked to epigenetics and disease[J].Trends Genet, 2001,17(9):520
  • 4Qi CF, Martensson A, Mattioli M, et al. CTCF functions as a critical regulator of cell-cycle arrest and death after ligation of the B cell receptor on immature B cells[J]. Proc Natl Acad Sci USA,2003,100(2):633
  • 5Fedoriw AM, Stein P, Svoboda P, Schultz, et al. Transgenic RNAi reveals essential function for CTCF in H19 gene imprinting[J]. Science, 2004,303(5655):238
  • 6Yusufzai TM, Tagami H, Nakatani Y, et al. CTCF tethers an insulator to subnuclear sites, suggesting shared insulator mechanisms across species[J]. Mol Cell, 2004,13(2):291
  • 7Chao W, Huynh KD, Spencer RJ, et al. CTCF, a candidate transacting factor for X-inactivation choice[J]. Science, 2002,295(5553):345
  • 8斯佩克特 戈德曼 莱因万德 著黄培堂.细胞实验指南(M)[M].北京:L科学出版社,2001.644-646.

同被引文献40

  • 1朱旭东,党颖,冯怡,李涛,黄培堂.用RNaseⅢ制备的小干涉RNA降解SARS冠状病毒基因的细胞内转录物[J].生物工程学报,2004,20(4):484-489. 被引量：5
  • 2杨煜,劳凤学,高振平,徐桂月,张鹏宇,柳忠辉.重组人FS315C末端蛋白表达及其抗体制备[J].中国生物工程杂志,2005,25(8):56-59. 被引量：2
  • 3郭威威,朱旭东,刘党生,黄培堂.腺相关病毒介导的RNA干扰抑制HeLa细胞中CTCF的表达[J].生物技术通讯,2006,17(4):503-505. 被引量：2
  • 4YEH A , WEI M , GOLUB S B , et al . Chromosome arm 16q in Wilms tumors: unbalanced chromosomal translocations, loss of heterozygosity, and assessment of the CTCF gene[J]. Genes Chromosomes. Cancer, 2002, 35 (2): 156- 163.
  • 5FILIPPOVA G N , LINDBLOM A , MEINCKE L J , et al.A widely expressed transcription factor with multiple DNA sequence specificity, CTCF, is localized at chromosome segment 16q22. 1 within one of the smallest regions of overlap for common deletions in breast and prostate cancers[J]. Genes Chromosomea. Cancer, 1998, 22(1): 26-36.
  • 6DARCI T , BUTCHERA D I , RODENHISER. Epigenetic inactivation of BRCA1 is associated with aberrant expression of CTCF and DNA methyltransferase (DNMT3B) in some sporadic breast tumours[J]. European Journal of Cancer, 2007, 43(1): 210-219.
  • 7DOCQUIER F , FARRAR D, D'ARCY V , et at . Heightened expression of CTCF in breast cancer cells is associated with resistance to apoptosis[J]. Cancer Res, 2005, 65(12): 5112-5122.
  • 8KLENOVA E M, NICOLAS R H, CARBE A F, et al. Molecular weight abnormalities of the CTCF transcription factor : CTCF migrates aberrantly in SDS-PAGE and the size of the expressed protein is affected by the UTEs and sequences within the coding region of the CTCF gene[J]. Nucleic Acids Res, 1997, 25(3): 466-474.
  • 9LOUKINOV D I , PUGACHEVA E , VATOLIN S , et al. BORIS, a novel male germ-line-specific protein associated with epigenetic reprogramming events, shares the same 11- zinc-finger domain with CTCF, the insulator protein involved in reading imprinting marks in the soma[J]. Proc Natl Acad Sci USA, 2002, 99(10): 6806-6811.
  • 10VOSTROV A A , TAHENY M J , QUITSCHKE W W. A region to the N-terminal side of the CTCF zinc finger domain is essential for activating transcription from the amyloid precursor protein[J]. J Biol Chem, 2002, 277(2): 1619-1627.

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生物技术通讯
2004年 第5期

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