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活跃的玉米基因未被修饰,其两侧是不同类别的经过修饰的高度重复DNA。

Active maize genes are unmodified and flanked by diverse classes of modified, highly repetitive DNA.

作者信息

Bennetzen J L, Schrick K, Springer P S, Brown W E, SanMiguel P

机构信息

Department of Biological Sciences, Purdue University, West Lafayette, IN 47907.

出版信息

Genome. 1994 Aug;37(4):565-76. doi: 10.1139/g94-081.

Abstract

We have characterized the copy number, organization, and genomic modification of DNA sequences within and flanking several maize genes. We found that highly repetitive DNA sequences were tightly linked to most of these genes. The highly repetitive sequences were not found within the coding regions but could be found within 6 kb either 3' or 5' to the structural genes. These highly repetitive regions were each composed of unique combinations of different short repetitive sequences. Highly repetitive DNA blocks were not interrupted by any detected single copy DNA. The 13 classes of highly repetitive DNA identified were found to vary little between diverse Zea isolates. The level of DNA methylation in and near these genes was determined by scoring the digestibility of 63 recognition/cleavage sites with restriction enzymes that were sensitive to 5-methylation of cytosines in the sequences 5'-CG-3' and 5'-CNG-3'. All but four of these sites were digestible in chromosomal DNA. The four undigested sites were localized to extragenic DNA within or near highly repetitive DNA, while the other 59 sites were in low copy number DNAs. Pulsed field gel analysis indicated that the majority of cytosine modified tracts range from 20 to 200 kb in size. Single copy sequences hybridized to the unmodified domains, while highly repetitive sequences hybridized to the modified regions. Middle repetitive sequences were found in both domains.

摘要

我们已经对几个玉米基因内部及其侧翼的DNA序列的拷贝数、组织形式和基因组修饰进行了表征。我们发现,高度重复的DNA序列与这些基因中的大多数紧密相连。在编码区域内未发现高度重复序列,但在结构基因的3'端或5'端6 kb范围内可以找到。这些高度重复区域各自由不同短重复序列的独特组合组成。高度重复的DNA块没有被任何检测到的单拷贝DNA打断。已发现所鉴定的13类高度重复DNA在不同的玉米分离株之间变化很小。通过对63个对序列5'-CG-3'和5'-CNG-3'中的胞嘧啶5-甲基化敏感的限制性内切酶识别/切割位点的消化率进行评分,确定了这些基因内部和附近的DNA甲基化水平。在染色体DNA中,除了四个位点外,所有这些位点都可被消化。四个未消化的位点定位于高度重复DNA内部或附近的基因外DNA,而其他59个位点位于低拷贝数DNA中。脉冲场凝胶分析表明,大多数胞嘧啶修饰片段的大小在20至200 kb之间。单拷贝序列与未修饰区域杂交,而高度重复序列与修饰区域杂交。在两个区域都发现了中度重复序列。

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