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黏粒衍生的和染色体特异性犬微卫星的应用。

Use of cosmid-derived and chromosome-specific canine microsatellites.

作者信息

Dickens H F, Holmes N G, Ryder E, Breen M, Thomas R, Suter N, Sampson J, Langford C F, Ross M, Carter N P, Binns M M

机构信息

Centre for Preventive Medicine, Animal Health Trust, Suffolk, England.

出版信息

J Hered. 1999 Jan-Feb;90(1):52-4. doi: 10.1093/jhered/90.1.52.

Abstract

The majority of microsatellite markers being used to generate the emerging genetic linkage maps of the dog are derived from small-insert, random clones. While such markers are easy to generate, they have the disadvantage that they cannot easily be physically mapped by fluorescence in situ hybridization (FISH), making it difficult to assess the extent of genome coverage represented by such maps. In contrast, microsatellite markers from large-insert libraries enable the linkage groups within which they fall to be physically anchored to specific chromosomes. One aim of our work is to identify at least one microsatellite-containing cosmid clone for each canine chromosome, to ensure that linkage groups exist for all chromosomes. This is particularly important for a species with as complex a karyotype as the dog. Locating two cosmids on each chromosome would allow the orientation of the linkage groups to be established. Chromosomal locations of cosmid clones containing microsatellites have been determined by FISH and confirmed using canine chromosome-specific paints. Microsatellite sequences have been genotyped on the DogMap reference family. Microsatellites derived from flow-sorted, chromosome-specific libraries represent another source of useful markers. Initial studies have been carried out on the canine X chromosome, on which markers were underrepresented in our initial studies.

摘要

用于构建犬类新出现的遗传连锁图谱的大多数微卫星标记,都来源于小插入片段的随机克隆。虽然这类标记易于生成,但它们存在一个缺点,即难以通过荧光原位杂交(FISH)进行物理定位,这使得评估此类图谱所代表的基因组覆盖范围变得困难。相比之下,来自大插入片段文库的微卫星标记能够将其所在的连锁群物理定位到特定染色体上。我们工作的一个目标是为每条犬染色体鉴定至少一个含微卫星的黏粒克隆,以确保所有染色体都存在连锁群。对于像犬这样核型复杂的物种来说,这一点尤为重要。在每条染色体上定位两个黏粒将有助于确定连锁群的方向。含微卫星的黏粒克隆的染色体位置已通过FISH确定,并使用犬染色体特异性探针进行了验证。微卫星序列已在DogMap参考家系中进行了基因分型。源自流式分选的染色体特异性文库的微卫星是另一种有用的标记来源。我们已经对犬X染色体进行了初步研究,在我们最初的研究中,该染色体上的标记较少。

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