Artieri C G, Mitchell L A, Ng S H S, Parisotto S E, Danzmann R G, Hoyheim B, Phillips R B, Morasch M, Koop B F, Davidson W S
Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada.
Cytogenet Genome Res. 2006;112(1-2):152-9. doi: 10.1159/000087528.
We have integrated data from linkage mapping, physical mapping and karyotyping to gain a better understanding of the sex-determining locus, SEX, in Atlantic salmon (Salmo salar). SEX has been mapped to Atlantic salmon linkage group 1 (ASL1) and is associated with several microsatellite markers. We have used probes designed from the flanking regions of these sex-linked microsatellite markers to screen a bacterial artificial chromosome (BAC) library, representing an 11.7x coverage of the Atlantic salmon genome, which has been HindIII fingerprinted and assembled into contigs. BACs containing sex-linked microsatellites and their related contigs have been identified and representative BACs have been placed on the Atlantic salmon chromosomes by fluorescent in situ hybridization (FISH). This identified chromosome 2, a large metacentric, as the sex chromosome. By positioning several BACs on this chromosome by FISH, it was possible to orient ASL1 with respect to chromosome 2. The region containing SEX appears to lie on the long arm between marker Ssa202DU and a region of heterochromatin identified by DAPI staining. BAC end-sequencing of clones within sex-linked contigs revealed five hitherto unmapped genes along the sex chromosome. We are using an in silico approach coupled with physical probing of the BAC library to extend the BAC contigs to provide a physical map of ASL1, with a view to sequencing chromosome 2 and, in the process, identifying the sex-determining gene.
我们整合了连锁图谱、物理图谱和核型分析的数据,以便更好地了解大西洋鲑(Salmo salar)的性别决定位点SEX。SEX已被定位到大西洋鲑连锁群1(ASL1),并与多个微卫星标记相关联。我们使用从这些性连锁微卫星标记侧翼区域设计的探针,筛选了一个细菌人工染色体(BAC)文库,该文库代表了大西洋鲑基因组11.7倍的覆盖率,已进行HindIII指纹图谱分析并组装成重叠群。已鉴定出含有性连锁微卫星及其相关重叠群的BAC,并通过荧光原位杂交(FISH)将代表性BAC定位到大西洋鲑染色体上。这确定了2号染色体,一条大型中着丝粒染色体,为性染色体。通过FISH将几个BAC定位到这条染色体上,得以确定ASL1相对于2号染色体的方向。包含SEX的区域似乎位于标记Ssa202DU和通过DAPI染色鉴定的异染色质区域之间的长臂上。对性连锁重叠群内克隆的BAC末端测序揭示了沿性染色体的五个迄今未定位的基因。我们正在使用计算机方法结合对BAC文库的物理探测来扩展BAC重叠群,以提供ASL1的物理图谱,以期对2号染色体进行测序,并在此过程中鉴定性别决定基因。