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犬Y染色体DNA序列:鉴定、测序及单核苷酸多态性发现

Dog Y chromosomal DNA sequence: identification, sequencing and SNP discovery.

作者信息

Natanaelsson Christian, Oskarsson Mattias C R, Angleby Helen, Lundeberg Joakim, Kirkness Ewen, Savolainen Peter

机构信息

School of Biotechnology, KTH, Royal Institute of Technology, AlbaNova University Center, 10691 Stockholm, Sweden.

出版信息

BMC Genet. 2006 Oct 6;7:45. doi: 10.1186/1471-2156-7-45.

Abstract

BACKGROUND

Population genetic studies of dogs have so far mainly been based on analysis of mitochondrial DNA, describing only the history of female dogs. To get a picture of the male history, as well as a second independent marker, there is a need for studies of biallelic Y-chromosome polymorphisms. However, there are no biallelic polymorphisms reported, and only 3200 bp of non-repetitive dog Y-chromosome sequence deposited in GenBank, necessitating the identification of dog Y chromosome sequence and the search for polymorphisms therein. The genome has been only partially sequenced for one male dog, disallowing mapping of the sequence into specific chromosomes. However, by comparing the male genome sequence to the complete female dog genome sequence, candidate Y-chromosome sequence may be identified by exclusion.

RESULTS

The male dog genome sequence was analysed by Blast search against the human genome to identify sequences with a best match to the human Y chromosome and to the female dog genome to identify those absent in the female genome. Candidate sequences were then tested for male specificity by PCR of five male and five female dogs. 32 sequences from the male genome, with a total length of 24 kbp, were identified as male specific, based on a match to the human Y chromosome, absence in the female dog genome and male specific PCR results. 14437 bp were then sequenced for 10 male dogs originating from Europe, Southwest Asia, Siberia, East Asia, Africa and America. Nine haplotypes were found, which were defined by 14 substitutions. The genetic distance between the haplotypes indicates that they originate from at least five wolf haplotypes. There was no obvious trend in the geographic distribution of the haplotypes.

CONCLUSION

We have identified 24159 bp of dog Y-chromosome sequence to be used for population genetic studies. We sequenced 14437 bp in a worldwide collection of dogs, identifying 14 SNPs for future SNP analyses, and giving a first description of the dog Y-chromosome phylogeny.

摘要

背景

迄今为止,对狗的群体遗传学研究主要基于线粒体DNA分析,仅描述了母犬的历史。为了了解公犬的历史以及获得第二个独立的标记,有必要研究双等位基因Y染色体多态性。然而,尚未有双等位基因多态性的报道,并且GenBank中仅存有3200 bp的非重复狗Y染色体序列,因此有必要鉴定狗Y染色体序列并寻找其中的多态性。目前仅对一只公犬的基因组进行了部分测序,无法将该序列定位到特定染色体上。但是,通过将公犬基因组序列与完整的母犬基因组序列进行比较,可以通过排除法鉴定出候选Y染色体序列。

结果

通过针对人类基因组的Blast搜索对公犬基因组序列进行分析,以鉴定与人类Y染色体匹配度最高的序列,并与母犬基因组进行比对以鉴定母犬基因组中不存在的序列。然后通过对5只公犬和5只母犬进行PCR,检测候选序列的雄性特异性。基于与人类Y染色体的匹配、在母犬基因组中的缺失以及雄性特异性PCR结果,从公犬基因组中鉴定出32个序列,总长度为24 kbp,具有雄性特异性。随后对来自欧洲、西南亚、西伯利亚、东亚、非洲和美洲的10只公犬的14437 bp进行了测序。发现了9种单倍型,由14个替换定义。单倍型之间的遗传距离表明它们至少起源于5种狼的单倍型。单倍型的地理分布没有明显趋势。

结论

我们已经鉴定出24159 bp的狗Y染色体序列用于群体遗传学研究。我们对来自世界各地的狗的14437 bp进行了测序,鉴定出14个单核苷酸多态性用于未来的SNP分析,并首次描述了狗Y染色体系统发育。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1bcd/1630699/f2625ed637f4/1471-2156-7-45-1.jpg

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