Figueroa Debbie M, Davis James D, Strobel Cornelia, Conejo Maria S, Beckham Katherine D, Ring Brian C, Bass Hank W
Department of Biological Science, The Florida State University, Tallahassee, FL 32306-4370, USA.
J Biomed Biotechnol. 2011;2011:386862. doi: 10.1155/2011/386862. Epub 2010 Dec 22.
The integration of genetic and physical maps of maize is progressing rapidly, but the cytogenetic maps lag behind, with the exception of the pachytene fluorescence in situ hybridization (FISH) maps of maize chromosome 9. We sought to produce integrated FISH maps of other maize chromosomes using Core Bin Marker loci. Because these 1 Kb restriction fragment length polymorphism (RFLP) probes are below the FISH detection limit, we used BACs from sorghum, a small-genome relative of maize, as surrogate clones for FISH mapping. We sequenced 151 maize RFLP probes and compared in silico BAC selection methods to that of library filter hybridization and found the latter to be the best. BAC library screening, clone verification, and single-clone selection criteria are presented along with an example of transgenomic BAC FISH mapping. This strategy has been used to facilitate the integration of RFLP and FISH maps in other large-genome species.
玉米遗传图谱与物理图谱的整合进展迅速,但细胞遗传图谱却相对滞后,不过玉米第9号染色体的粗线期荧光原位杂交(FISH)图谱除外。我们试图利用核心库位标记基因座制作其他玉米染色体的整合FISH图谱。由于这些1千碱基限制性片段长度多态性(RFLP)探针低于FISH检测极限,我们使用高粱(玉米的小基因组近缘物种)的细菌人工染色体(BAC)作为FISH定位的替代克隆。我们对151个玉米RFLP探针进行了测序,并将计算机模拟BAC选择方法与文库滤膜杂交法进行了比较,发现后者是最佳方法。文中介绍了BAC文库筛选、克隆验证和单克隆选择标准,并给出了转基因BAC FISH定位的实例。该策略已被用于促进其他大基因组物种中RFLP图谱与FISH图谱的整合。