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一组99个牛微卫星:特征分析、同线定位及多态性研究

A set of 99 cattle microsatellites: characterization, synteny mapping, and polymorphism.

作者信息

Vaiman D, Mercier D, Moazami-Goudarzi K, Eggen A, Ciampolini R, Lépingle A, Velmala R, Kaukinen J, Varvio S L, Martin P

机构信息

Laboratoire de Génétique biochimique et de Cytogénétique, INRA-CRJ, Jouy-en-Josas, France.

出版信息

Mamm Genome. 1994 May;5(5):288-97. doi: 10.1007/BF00389543.

Abstract

Cattle microsatellite clones (136) were isolated from cosmid (10) and plasmid (126) libraries and sequenced. The dinucleotide repeats were studied in each of these sequences and compared with dinucleotide repeats found in other vertebrate species where information was available. The distribution in cattle was similar to that described for other mammals, such as rat, mouse, pig, or human. A major difference resides in the number of sequences present in the bovine genome, which seemed at best one-third as large as in other species. Oligonucleotide primers (117 pairs) were synthesized, and a PCR product of expected size was obtained for 88 microsatellite sequences (75%). Synteny or chromosome assignment was searched for each locus with PCR amplification on a panel of 36 hamster/bovine somatic cell hybrids. Of our bovine microsatellites, eighty-six could be assigned to synteny groups of chromosomes. In addition, 10 other microsatellites--HEL 5, 6, 9, 11, 12, 13 (Kaukinen and Varvio 1993), HEL 4, 7, 14, 15--as well as the microsatellite found in the kappa-casein gene (Fries et al. 1990) were mapped on the hybrids. Microsatellite polymorphism was checked on at least 30 unrelated animals of different breeds. Almost all the autosomal and X Chr microsatellites displayed polymorphism, with the number of alleles varying between two and 44. We assume that these microsatellites could be very helpful in the construction of a primary public linkage map of the bovine genome, with an aim of finding markers for Economic Trait Loci (ETL) in cattle.

摘要

从黏粒文库(10个)和质粒文库(126个)中分离出136个牛微卫星克隆并进行测序。对这些序列中的每一个二核苷酸重复序列进行了研究,并与其他有信息可查的脊椎动物物种中发现的二核苷酸重复序列进行了比较。牛中二核苷酸重复序列的分布与其他哺乳动物(如大鼠、小鼠、猪或人类)中描述的相似。一个主要差异在于牛基因组中存在的序列数量,其似乎最多只有其他物种的三分之一。合成了117对寡核苷酸引物,88个微卫星序列(75%)获得了预期大小的PCR产物。利用一组36个仓鼠/牛体细胞杂种进行PCR扩增,为每个位点寻找同线性或染色体定位。在我们的牛微卫星中,86个可以被定位到染色体同线性组。此外,另外10个微卫星——HEL 5、6、9、11、12、13(考基宁和瓦尔维奥,1993年)、HEL 4、7、14、15——以及κ-酪蛋白基因中发现的微卫星(弗里斯等人,1990年)被定位到杂种细胞上。在至少30只不同品种的无亲缘关系动物中检测了微卫星多态性。几乎所有常染色体和X染色体微卫星都显示出多态性,等位基因数量在2到44之间变化。我们认为这些微卫星在构建牛基因组初级公共连锁图谱方面可能非常有用,目的是寻找牛经济性状基因座(ETL)的标记。

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