Mungall A J, Edwards C A, Ranby S A, Humphray S J, Heathcott R W, Clee C M, East C L, Holloway E, Butler A P, Langford C F, Gwilliam R, Rice K M, Maslen G L, Carter N P, Ross M T, Deloukas P, Bentley D R, Dunham I
Sanger Centre, Huxton, Cambridge, UK.
DNA Seq. 1996;7(1):47-9. doi: 10.3109/10425179609015647.
The development of radiation hybrid (RH) mapping (Cox et al., 1990) and the availability of large numbers of STS markers, together with extensive bacterial clone resources provided a means to accelerate the process of mapping a human chromosome and preparing bacterial clone contigs ready to sequence. Our aim is to construct physical clone maps covering those regions of chromosome 6 that are not currently extensively mapped, and use these to determine the DNA sequence of the whole chromosome. We report here a strategy which initially involves establishing a high density framework map using RH mapping. The framework markers are then used for the identification of bacterial genomic clones covering the chromosome. The bacterial clones are analysed by restriction enzyme fingerprinting and STS-content analysis to identify sequence-ready contigs. Contig gap closure will also be performed by clone walking.
辐射杂种(RH)图谱绘制技术的发展(考克斯等人,1990年)以及大量序列标签位点(STS)标记的可得性,再加上丰富的细菌克隆资源,为加速人类染色体图谱绘制进程以及制备可供测序的细菌克隆重叠群提供了一种手段。我们的目标是构建覆盖6号染色体上目前尚未广泛绘制区域的物理克隆图谱,并利用这些图谱确定整条染色体的DNA序列。我们在此报告一种策略,该策略首先涉及利用RH图谱绘制技术建立高密度框架图谱。然后,框架标记用于鉴定覆盖该染色体的细菌基因组克隆。通过限制性酶切指纹分析和STS含量分析对细菌克隆进行分析,以鉴定可供测序的重叠群。重叠群的缺口填补也将通过克隆步移来完成。