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利用双等位基因标记对拟南芥进行全基因组图谱绘制。

Genome-wide mapping with biallelic markers in Arabidopsis thaliana.

作者信息

Cho R J, Mindrinos M, Richards D R, Sapolsky R J, Anderson M, Drenkard E, Dewdney J, Reuber T L, Stammers M, Federspiel N, Theologis A, Yang W H, Hubbell E, Au M, Chung E Y, Lashkari D, Lemieux B, Dean C, Lipshutz R J, Ausubel F M, Davis R W, Oefner P J

机构信息

Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA.

出版信息

Nat Genet. 1999 Oct;23(2):203-7. doi: 10.1038/13833.

Abstract

Single-nucleotide polymorphisms, as well as small insertions and deletions (here referred to collectively as simple nucleotide polymorphisms, or SNPs), comprise the largest set of sequence variants in most organisms. Positional cloning based on SNPs may accelerate the identification of human disease traits and a range of biologically informative mutations. The recent application of high-density oligonucleotide arrays to allele identification has made it feasible to genotype thousands of biallelic SNPs in a single experiment. It has yet to be established, however, whether SNP detection using oligonucleotide arrays can be used to accelerate the mapping of traits in diploid genomes. The cruciferous weed Arabidopsis thaliana is an attractive model system for the construction and use of biallelic SNP maps. Although important biological processes ranging from fertilization and cell fate determination to disease resistance have been modelled in A. thaliana, identifying mutations in this organism has been impeded by the lack of a high-density genetic map consisting of easily genotyped DNA markers. We report here the construction of a biallelic genetic map in A. thaliana with a resolution of 3.5 cM and its use in mapping Eds16, a gene involved in the defence response to the fungal pathogen Erysiphe orontii. Mapping of this trait involved the high-throughput generation of meiotic maps of F2 individuals using high-density oligonucleotide probe array-based genotyping. We developed a software package called InterMap and used it to automatically delimit Eds16 to a 7-cM interval on chromosome 1. These results are the first demonstration of biallelic mapping in diploid genomes and establish means for generalizing SNP-based maps to virtually any genetic organism.

摘要

单核苷酸多态性以及小的插入和缺失(在此统称为单核苷酸多态性,即SNPs)构成了大多数生物体中最大的序列变异集合。基于SNPs的定位克隆可能会加速人类疾病性状和一系列具有生物学信息的突变的鉴定。最近将高密度寡核苷酸阵列应用于等位基因鉴定,使得在单个实验中对数千个双等位基因SNPs进行基因分型成为可能。然而,使用寡核苷酸阵列进行SNP检测是否可用于加速二倍体基因组中性状的定位,目前尚未确定。十字花科杂草拟南芥是构建和使用双等位基因SNP图谱的一个有吸引力的模型系统。尽管从受精、细胞命运决定到抗病性等重要生物学过程已在拟南芥中得到模拟,但由于缺乏由易于进行基因分型的DNA标记组成的高密度遗传图谱,在该生物体中鉴定突变受到了阻碍。我们在此报告了在拟南芥中构建分辨率为3.5 cM的双等位基因遗传图谱及其在定位Eds16中的应用,Eds16是一个参与对真菌病原体白粉菌防御反应的基因。该性状的定位涉及使用基于高密度寡核苷酸探针阵列基因分型的F2个体减数分裂图谱的高通量生成。我们开发了一个名为InterMap的软件包,并使用它将Eds16自动定位到第1号染色体上一个7-cM的区间。这些结果首次证明了在二倍体基因组中的双等位基因定位,并建立了将基于SNP的图谱推广到几乎任何遗传生物体的方法。

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