Yokota H, Takahashi M, Iwasaki T, Oishi M
Institute of Applied Microbiology, University of Tokyo, Japan.
Cell Struct Funct. 1989 Dec;14(6):759-67. doi: 10.1247/csf.14.759.
By employing in gel competitive reassociation, which distinguishes two DNA preparations, to clone anonymous DNA fragments with altered primary structure, we isolated a clone (BL-1) from a rat DNA which gave an extra, apparently altered, DNA band in a specific tissue (brain) when we used it as a probe for Southern hybridization. The sequence of BL-1 was very similar to a portion of LINE, a highly repetitive sequence. Southern hybridization analysis with oligonucleotide probes representing a portion of the BL-1 sequence confirmed the presence of the brain-specific band. Another, different band specific to HindIII digests of heart DNA was also detected with the same oligonucleotide probes. Furthermore, we found that poly (A) +RNA hybridized with BL-1 was enriched in brain. These results suggest that there are tissue specific alteration of genomic DNA in rats which may be associated with transcriptional activation specific to the sequence. Alternative explanations of the results are also discussed.
通过采用凝胶内竞争性复性(可区分两种DNA制剂)来克隆具有改变的一级结构的匿名DNA片段,我们从大鼠DNA中分离出一个克隆(BL-1),当我们将其用作Southern杂交探针时,它在特定组织(脑)中产生了一条额外的、明显改变的DNA条带。BL-1的序列与LINE的一部分非常相似,LINE是一种高度重复序列。用代表BL-1序列一部分的寡核苷酸探针进行Southern杂交分析证实了脑特异性条带的存在。用相同的寡核苷酸探针还检测到心脏DNA的HindIII消化物特有的另一条不同的条带。此外,我们发现与BL-1杂交的聚(A)+RNA在脑中富集。这些结果表明,大鼠基因组DNA存在组织特异性改变,这可能与该序列特异性的转录激活有关。还讨论了对结果的其他解释。