Barbosa Angela, Demeure Olivier, Urien Céline, Milan Denis, Chardon Patrick, Renard Christine
Laboratoire mixte de Radiobiologie et d'Etude du Génome, Institut National de la Recherche Agronomigue et Center d'Energie Atomique, Domaine de Vilvert, 78352, Jouy en Josas cedex, France.
Mamm Genome. 2004 Dec;15(12):982-95. doi: 10.1007/s00335-004-3008-6.
The aim of this study was to establish a porcine physical map along the chromosome SSC7q by construction of BAC contigs between microsatellites Sw1409 and S0102. The SLA class II contig, located on SSC7q, was lengthened. Four major BAC contigs and 10 short contigs span a region equivalent to 800 cR measured by IMpRH7000 mapping. The BAC contigs were initiated by PCR screening with primers derived from human orthologous segments, extended by chromosome walking, and controlled and oriented by RH mapping with the two available panels, IMpRH7000Rad and IMNpRH12000Rad. The location of 43 genes was revealed by sequenced segments, either from BAC ends or PCR products from BAC clones. The 220 BAC end sequences (BES) were also used to analyze the different marks of evolution. Comparative mapping analysis between pigs and humans demonstrated that the gene organization on HSA6p21 and on SSC7p11 and q11-q14 segments was conserved during evolution, with the exception of long fragments of HSA6p12 which shuffled and spliced the SLA extended class II region. Additional punctual variations (unique gene insertion/deletion) were observed, even within conserved segments, revealing the evolutionary complexity of this region. In addition, 18 new polymorphic microsatellites have been selected in order to cover the entire SSC7p11-q14 region.
本研究的目的是通过构建微卫星Sw1409和S0102之间的细菌人工染色体(BAC)重叠群,建立一条沿猪7号染色体长臂(SSC7q)的物理图谱。位于SSC7q上的SLA II类重叠群得以延长。四个主要的BAC重叠群和10个短重叠群跨越了一个相当于通过IMpRH7000图谱测量的800厘摩(cR)的区域。BAC重叠群通过用源自人类同源片段的引物进行PCR筛选来起始,通过染色体步移来延伸,并通过使用两个可用的辐射杂种板IMpRH7000Rad和IMNpRH12000Rad进行辐射杂种图谱分析来控制和定向。通过测序片段(来自BAC末端或BAC克隆的PCR产物)揭示了43个基因的位置。220个BAC末端序列(BES)也用于分析不同的进化标记。猪和人类之间的比较图谱分析表明,除了人类6号染色体短臂(HSA6p)12的长片段对SLA扩展II类区域进行了重排和拼接外,HSA6p21以及SSC7p11和q11 - q14区段上的基因组织在进化过程中是保守的。即使在保守区段内也观察到了额外的点状变异(独特的基因插入/缺失),揭示了该区域进化的复杂性。此外,为了覆盖整个SSC7p11 - q14区域,已选择了18个新的多态微卫星。