Tissue and strain-specific patterns of endogenous proviral hypomethylation analyzed by two-dimensional gel electrophoresis.
Tissue and strain-specific patterns of endogenous proviral hypomethylation analyzed by two-dimensional gel electrophoresis.
复制标题
通过二维凝胶电泳分析内源性前病毒低甲基化的组织和菌株特异性模式。
DOI：
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发表时间：
1990
影响因子：
11.1
通讯作者：
E. Kuff
中科院分区：
文献类型：
--
作者：
J. Mietz;E. Kuff
与脑池内A颗粒（IAP）相关的前病毒序列被扩增并分散在小鼠基因组中。它们的表达与5'长末端重复序列中CpG位点的低甲基化有关。我们已经使用二维琼脂糖凝胶电泳检测小鼠DNA中IAP低甲基化的模式。该方法对5'长末端重复序列中保守Hae II位点的甲基化状态和每个低甲基化长末端重复序列上游侧翼DNA中最近的BamHI位点的位置都敏感。该方法还检测来自IAP元件的限制性片段，所述IAP元件本身是甲基化的，但在其5'相邻DNA中具有未甲基化的Hae II位点。从四个近交系小鼠品系（BALB/c，C3 H/He，C57 BL/6，和DBA/2）的DNA给出了独特的二维模式的BamHI/Hae II限制性片段与IAP探针杂交检测。这种组成模式在很大程度上是保守的几个组织中的每个菌株，但观察到一些组织特异性的变化。在二维模式中反映的位点特异性低甲基化是可遗传的特性，因为来自株间杂交后代的DNA包含两组亲本片段。IAP元件可能是有用的基因组甲基化模式的内源性报告基因。
Proviral sequences related to the intracisternal A particle (IAP) are amplified and dispersed in the mouse genome. Their expression is associated with hypomethylation at CpG sites in the 5' long terminal repeat. We have used two-dimensional agarose gel electrophoresis to examine patterns of IAP hypomethylation in mouse DNA. The method is sensitive to both the methylation status of a conserved Hae II site in the 5' long terminal repeat and the location of the closest BamHI site in the flanking DNA upstream of each hypomethylated long terminal repeat. The method also defects restriction fragments derived from IAP elements that are themselves methylated but have an unmethylated Hae II site in their 5' adjacent DNA. DNAs from each of four inbred mouse strains (BALB/c, C3H/He, C57BL/6, and DBA/2) gave distinctive two-dimensional patterns of BamHI/Hae II restriction fragments detected by hybridization with an IAP probe. This constitutive pattern was largely conserved among several tissues of each strain, but some tissue-specific variations were observed. The site-specific hypomethylations reflected in the two-dimensional patterns were heritable properties, since DNA from progeny of an interstrain cross contained both parental sets of fragments. IAP elements may be useful endogenous reporters of genomic methylation patterns.