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马铃薯简单序列重复位点的分离、特征分析及定位

Isolation, characterisation and mapping of simple sequence repeat loci in potato.

作者信息

Milbourne D, Meyer R C, Collins A J, Ramsay L D, Gebhardt C, Waugh R

机构信息

Department of Cell and Molecular Genetics, Scottish Crop Research Institute, Invergowrie, Dundee, UK.

出版信息

Mol Gen Genet. 1998 Aug;259(3):233-45. doi: 10.1007/s004380050809.

Abstract

Solanum tuberosum L. DNA sequences containing simple sequence repeat (SSR) motifs were extracted from the EMBL database, cDNA and selectively enriched small-insert DNA libraries. Enrichment was achieved using either triplex affinity capture or single-strand hybridisation selection. One hundred and twelve primer pairs which successfully amplified products of the correct size from potato DNA were ultimately designed and synthesised. Ninety-eight of these revealed length polymorphisms in a panel of four diploid and two tetraploid clones, in agreement with the high information content of this class of markers which has been found in other species. All of the markers were assigned a quality score of 1-5 based on their potential usefulness. Eighty-nine loci from 65 of the primer pairs were located on two genetic linkage maps of potato by segregation analysis of the amplified alleles. Fifty-two of the SSRs were clearly single locus. The maps were aligned using 23 SSR primer pairs and 13 RFLP loci mapped in both populations. The markers described constitute a class which should replace Restriction Fragment Length Polymorphisms (RFLP) as the markers of choice for future genetic studies in potato. The sequences of the primers, together with other information on these markers are provided.

摘要

从EMBL数据库、cDNA以及选择性富集的小插入DNA文库中提取了含有简单序列重复(SSR)基序的马铃薯(Solanum tuberosum L.)DNA序列。富集是通过三链亲和捕获或单链杂交选择实现的。最终设计并合成了112对引物,这些引物能成功地从马铃薯DNA中扩增出正确大小的产物。其中98对引物在一组4个二倍体和2个四倍体克隆中显示出长度多态性,这与在其他物种中发现的此类标记的高信息含量一致。根据其潜在用途,所有标记都被赋予了1 - 5的质量评分。通过对扩增等位基因的分离分析,来自65对引物的89个位点定位在马铃薯的两个遗传连锁图谱上。其中52个SSR明显是单一位点。利用在两个群体中都定位的23对SSR引物和13个RFLP位点对图谱进行了比对。所描述的这些标记构成了一类标记,应该取代限制性片段长度多态性(RFLP),成为未来马铃薯遗传研究的首选标记。文中提供了引物序列以及关于这些标记的其他信息。

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