He L, Zhu Z, Faras A J, Guise K S, Hackett P B, Kapuscinski A R
Department of Fisheries and Wildlife, University of Minnesota, St Paul 55108.
Mol Mar Biol Biotechnol. 1992 Apr;1(2):125-35.
Two families of repetitive DNA sequences were isolated from the zebrafish genome and characterized. Eight different sequences were sequenced and classified by two standards, their (G + C) composition and their lengths. For convenience, the sequences were first divided into two types. Type I was (A + T)-rich, was repeated approximately 500,000 times, and constituted approximately 5% of the zebrafish genome. Type II was (G + C)-rich, was reiterated approximately 90,000 times, and comprised approximately 0.5% of the genome. Agarose gel electrophoresis of zebrafish DNA cleaved with AluI revealed three distinguishable bands of repetitive fragments: large (approximately 180 bp, designated RFAL), medium (approximately 140 bp, RFAM), and small (approximately 90 bp, RFAS). The RFAL fragments contained both type I and type II sequences. Limited digestion of genomic DNA indicated that RFAL and RFAM were tandemly arranged in the genome, whereas RFAS showed a mixed pattern of both tandem and interspersed repeated arrangements. Although inclusion of a repetitive sequence in a transgenic construct did not appreciably accelerate homologous integration of transgenes into the zebrafish genome, the AluI sequences could facilitate transgene mapping following chromosomal integration.
从斑马鱼基因组中分离并鉴定了两个重复DNA序列家族。对八个不同的序列按照两个标准进行了测序和分类,即它们的(G + C)组成和长度。为方便起见,这些序列首先被分为两种类型。I型富含(A + T),重复约500,000次,约占斑马鱼基因组的5%。II型富含(G + C),重复约90,000次,约占基因组的0.5%。用AluI切割的斑马鱼DNA进行琼脂糖凝胶电泳,显示出三条可区分的重复片段带:大的(约180 bp,命名为RFAL)、中等的(约140 bp,RFAM)和小的(约90 bp,RFAS)。RFAL片段包含I型和II型序列。对基因组DNA的有限消化表明,RFAL和RFAM在基因组中是串联排列的,而RFAS则呈现出串联和散布重复排列的混合模式。尽管在转基因构建体中包含重复序列并没有明显加速转基因在斑马鱼基因组中的同源整合,但AluI序列可以在染色体整合后促进转基因的定位。