An arrayed human not I-EcoRV boundary library as a tool for RLGS spot analysis.
An arrayed human not I-EcoRV boundary library as a tool for RLGS spot analysis.
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阵列人类非 I-EcoRV 边界库作为 RLGS 点分析的工具。
DOI：
10.1093/dnares/4.3.253
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Held,WA
中科院分区：
文献类型：
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作者：
Plass,C;Weichenhan,D;Catanese,J;Costello,JF;Yu,F;Yu,L;Smiraglia,D;Cavenee,WK;Caligiuri,MA;deJong,P;Held,WA
限制性标记基因组扫描（RLGS）是一种允许检测两个DNA样品之间的遗传差异的方法。它是基于罕见的切割限制性位点的末端标记和通过二维凝胶电泳分离这些片段。1RLGS图谱显示超过2000个片段或”斑点”，其强度与特定序列的拷贝数相关。RLGS中使用的大多数稀有切割限制性内切酶具有富含GC的识别序列，其主要存在于哺乳动物基因组中的CpG岛中。CpG岛位于所有已知管家基因和40%组织特异性基因的5 '调控区。2'3因此，RLGS图谱强烈偏向于显示位于基因附近的序列，而不是随机的基因组DNA片段。在人类中，RLGS已被几个小组用于检测伴随人类肿瘤发生的遗传和表观遗传变化。4'5 RLGS分析的一个基本步骤是克隆对应于RLGS斑点的DNA序列，以鉴定受特异性改变影响的基因。已经描述了几种RLGS斑点克隆程序，并且基于基因组DNA的限制性捕获纯化以减少未标记的背景片段的量，6或涉及接头序列与斑点DNA末端的连接，随后进行聚合酶链式反应（PCR），7或两种方法的组合。5然而，RLGS斑点克隆的大小
Restriction landmark genomic scanning (RLGS) is a method which allows the detection of genetic differences between two DNA samples. It is based on the endlabeling of rare cutting restriction sites and the separation of these fragments by two-dimensional gel electrophoresis. 1 RLGS profiles display over 2000 fragments, or" spots," having intensities which correlate with the copy number of the particular sequence. Most of the rare cutting restriction enzymes used in RLGS possess GC-rich recognition sequences that are mainly found in CpG-islands in mammalian genomes. CpG islands are located in the 5'regulatory regions of all known housekeeping genes and 40% of tissue-specific genes. 2'3 Thus, RLGS profiles are strongly biased towards the display of sequences located in the vicinity of genes, rather than random genomic DNA fragments. In humans, RLGS has been used by several groups for the detection of genetic and epigenetic changes which accompany human tumorigenesis. 4'5 An essential step in RLGS analysis is the cloning of DNA sequences corresponding to RLGS spots in order to identify genes affected by the specific alteration. Several RLGS spot cloning procedures have been described and are based on either a restriction trapper purification of genomic DNA to reduce the amount of unlabeled background fragments, 6 or involve the ligation of adaptor sequences to the ends of the spot DNA followed by polymerase chain reaction (PCR), 7 or a combination of the two methods. 5 However, the size of RLGS spot clones