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微卫星 DNA 标记的开发及其在普通狨猴中的染色体定位。

Development of microsatellite DNA markers and their chromosome assignment in the common marmoset.

机构信息

Institute for Experimental Animals, Hamamatsu University School of Medicine, Hamamatsu, Japan.

出版信息

Am J Primatol. 2009 Nov;71(11):912-8. doi: 10.1002/ajp.20729.

DOI:10.1002/ajp.20729
PMID:19637280
Abstract

This study was performed to develop microsatellite DNA markers, which are useful for linkage analyses, gene mapping and blood chimera analyses in the common marmoset (Callithrix jacchus). We searched 153 of 295 bacterial artificial clone DNA sequences of the common marmoset that were archived in the NCBI database in 2004. On the basis of the search, we designed 186 PCR primer sets. When tested using 5 unrelated individuals, we successfully detected 154 markers with PCR products, of which 80 (52%) were polymorphic and 74 (48%) were monomorphic. We assigned each of the 154 markers to a human chromosome based on BLAST searches, which was achieved by searching the entire human genome sequences using an approximately 3 kb section of each forward primer sequence, including approximately 1.5 kb of the upstream and approximately 1.5 kb of the downstream sequences. Combining our assignment data and the chromosome painting-assisted karyotype of the common marmoset [Sherlock et al., Genomics 33:214-219, 1996], we prepared a list of 154 microsatellite DNA markers that were assigned to human chromosomes, except for the Y chromosome, which is equivalent to a chromosome map. Using five microsatellite DNA markers, we have established a fragment analysis method with a sequencer, which can be routinely used for blood chimera analysis, parentage diagnosis and individual identification.

摘要

本研究旨在开发微卫星 DNA 标记物,这些标记物可用于普通狨猴(Callithrix jacchus)的连锁分析、基因定位和血液嵌合体分析。我们在 2004 年对 NCBI 数据库中归档的普通狨猴 295 个细菌人工克隆 DNA 序列中的 153 个进行了搜索。在此基础上,我们设计了 186 对 PCR 引物。在使用 5 个无关个体进行测试时,我们成功地检测到了 154 个具有 PCR 产物的标记物,其中 80 个(52%)为多态性,74 个(48%)为单态性。我们根据 BLAST 搜索将这 154 个标记物中的每一个分配到人类染色体上,这是通过使用每个正向引物序列的大约 3kb 部分,包括大约 1.5kb 的上游和大约 1.5kb 的下游序列,对整个人类基因组序列进行搜索来实现的。结合我们的分配数据和普通狨猴的染色体作图辅助核型[Sherlock 等人,Genomics 33:214-219,1996],我们准备了一份 154 个微卫星 DNA 标记物的列表,这些标记物被分配到人类染色体上,除了 Y 染色体,它相当于染色体图谱。我们使用五个微卫星 DNA 标记物,建立了一种使用测序仪进行片段分析的方法,该方法可常规用于血液嵌合体分析、亲子关系诊断和个体识别。

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Primates. 2015 Jul;56(3):235-40. doi: 10.1007/s10329-015-0466-2. Epub 2015 Mar 25.