Allele-specific analysis of transcription factors binding to promoter regions
Allele-specific analysis of transcription factors binding to promoter regions
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DOI：
10.1016/s1046-2023(02)00004-x
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发表时间：
2002-01-01
期刊：
影响因子：
4.8
通讯作者：
Boxer, LM
中科院分区：
文献类型：
--
作者：
Heckman, CA;Boxer, LM
体内足迹技术可用于鉴定介导基因转录控制的调控元件。然而，基因的调控在两个等位基因之间可以不同，并且必须采取进一步的步骤来区分在一个等位基因上占据的调控元件和在第二个等位基因上使用的调控元件。许多血液恶性肿瘤是由染色体易位引起的，在某些情况下，染色体易位将基因重新定位到转录活性区域，导致该基因的表达失调。这种情况提供了两个等位基因之间差异表达的实例。在研究T（14; 18）和t（8; 14）易位，分别涉及bcl-2和c-myc原oneogenes，我们已经能够鉴定在介导易位等位基因的激活和正常等位基因的沉默中重要的调节元件。在体内甲基化和分离基因组DNA后，我们能够通过电泳分离易位和正常等位基因。使用连接介导的聚合酶链反应（LMPCR）技术，我们可以评估两个不同等位基因上的蛋白质相互作用。这种方法的详细描述与我们的研究中的例子提供了讨论这些技术如何可以应用到其他基因的研究。(C)2002 Elsevier Science（美国）。All rights reserved.
In vivo footprinting techniques are useful for the identification of regulatory elements mediating transcriptional control of a gene. However, regulation of a gene can differ between the two alleles, and further steps must be taken to distinguish between the regulatory elements occupied on one allele and those used on the second allele. Many hematologic malignancies result from chromosomal translocations, which, in some cases, relocate a gene to a transcriptionally active region leading to the deregulated expression of that gene. This situation provides an example of differential expression between two alleles. In studying the t(14; 18) and t(8; 14) translocations, which involve the bcl-2 and c-myc proto-oneogenes, respectively, we have been able to identify regulatory elements important in mediating the activation of the translocated alleles and the silencing of the normal alleles. Following in vivo methylation and isolation of genomic DNA, we were able to separate the translocated and normal alleles by electrophoresis. Using the ligation-mediated polyrnerase chain reaction (LMPCR) technique, we could then assess protein interactions on the two different alleles. A detailed description of this methodology with examples from our studies are provided with a discussion of how these techniques may be applied to the study of other genes. (C) 2002 Elsevier Science (USA). All rights reserved.