Raudsepp Terje, Santani Avni, Wallner Barbara, Kata Srinivas R, Ren Chengwei, Zhang Hong-Bin, Womack James E, Skow Loren C, Chowdhary Bhanu P
Department of Veterinary Anatomy and Public Health, Texas A&M University, College Station, 77843, USA.
Proc Natl Acad Sci U S A. 2004 Jun 22;101(25):9321-6. doi: 10.1073/pnas.0403011101. Epub 2004 Jun 14.
We herein report a detailed physical map of the horse Y chromosome. The euchromatic region of the chromosome comprises approximately 15 megabases (Mb) of the total 45- to 50-Mb size and lies in the distal one-third of the long arm, where the pseudoautosomal region (PAR) is located terminally. The rest of the chromosome is predominantly heterochromatic. Because of the unusual organization of the chromosome (common to all mammalian Y chromosomes), a number of approaches were used to crossvalidate the results. Analysis of the 5,000-rad horse x hamster radiation hybrid panel produced a map spanning 88 centirays with 8 genes and 15 sequence-tagged site (STS) markers. The map was verified by several fluorescence in situ hybridization approaches. Isolation of bacterial artificial chromosome (BAC) clones for the radiation hybrid-mapped markers, end sequencing of the BACs, STS development, and bidirectional chromosome walking yielded 109 markers (100 STS and 9 genes) contained in 73 BACs. STS content mapping grouped the BACs into seven physically ordered contigs (of which one is predominantly ampliconic) that were verified by metaphase-, interphase-, and fiber-fluorescence in situ hybridization and also BAC fingerprinting. The map spans almost the entire euchromatic region of the chromosome, of which 20-25% (approximately 4 Mb) is covered by isolated BACs. The map is presently the most informative among Y chromosome maps in domesticated species, third only to the human and mouse maps. The foundation laid through the map will be critical in obtaining complete sequence of the euchromatic region of the horse Y chromosome, with an aim to identify Y specific factors governing male infertility and phenotypic sex variation.
我们在此报告马Y染色体的详细物理图谱。该染色体的常染色质区域在总长45至50兆碱基(Mb)中约占15兆碱基,位于长臂的远侧三分之一处,假常染色体区域(PAR)位于其末端。染色体的其余部分主要是异染色质。由于该染色体的特殊组织形式(所有哺乳动物Y染色体均如此),我们采用了多种方法来交叉验证结果。对5000拉德的马×仓鼠辐射杂种细胞系进行分析,构建了一个跨越88厘雷的图谱,其中包含8个基因和15个序列标签位点(STS)标记。该图谱通过多种荧光原位杂交方法进行了验证。通过分离辐射杂种图谱标记的细菌人工染色体(BAC)克隆、BAC末端测序、STS开发以及双向染色体步移,获得了包含在73个BAC中的109个标记(100个STS和9个基因)。STS含量图谱将BAC分为7个物理有序的重叠群(其中一个主要是扩增子性),这些重叠群通过中期、间期和纤维荧光原位杂交以及BAC指纹图谱得到了验证。该图谱几乎覆盖了染色体的整个常染色质区域,其中20%至25%(约4兆碱基)被分离的BAC覆盖。目前,该图谱是家养物种Y染色体图谱中信息最丰富的,仅次于人类和小鼠图谱,位列第三。通过该图谱奠定基础对于获得马Y染色体常染色质区域的完整序列至关重要,目的是识别控制雄性不育和表型性别变异的Y特异性因子。