Osoegawa Kazutoyo, Zhu Baoli, Shu Chung Li, Ren Teresa, Cao Qing, Vessere Gery M, Lutz Michelle M, Jensen-Seaman Michael I, Zhao Shaying, de Jong Pieter J
Children's Hospital and Research Center at Oakland, Oakland, California 94609, USA.
Genome Res. 2004 Apr;14(4):780-5. doi: 10.1101/gr.2033904.
Two 11-fold redundant bacterial artificial chromosome (BAC) libraries (RPCI-32 and CHORI-230) have been constructed to support the rat genome project. The first library was constructed using a male Brown Norway (BN/SsNHsd) rat as a DNA source long before plans for rat genome sequencing had been launched. The second library was prepared from a highly inbred female (BN/SsNHsd/MCW) rat in support of the rat genome sequencing project. The use of an inbred rat strain is essential to avoid problems with genome assembly resulting from the difficulty of distinguishing haplotype variation from variation among duplicons. We have demonstrated the suitability of the library by using a detailed quality assessment of large insert sizes, narrow size distribution, consistent redundancy for many markers, and long-range continuity of BAC contig maps. The widespread use of the two libraries as an integral part of the rat genome project has led to the database annotations for many clones, providing rat researchers with a rich resource of BAC clones that can be screened in silico for genes of interest.
为支持大鼠基因组计划,已构建了两个具有11倍冗余度的细菌人工染色体(BAC)文库(RPCI - 32和CHORI - 230)。第一个文库早在大鼠基因组测序计划启动之前,就以雄性挪威棕鼠(BN/SsNHsd)作为DNA来源构建而成。第二个文库则是由一只高度近交的雌性(BN/SsNHsd/MCW)大鼠制备,以支持大鼠基因组测序项目。使用近交大鼠品系对于避免因难以区分单倍型变异和重复子间变异而导致的基因组组装问题至关重要。我们通过对大插入片段大小进行详细的质量评估、窄大小分布、许多标记的一致冗余度以及BAC重叠群图谱的长程连续性,证明了该文库的适用性。这两个文库作为大鼠基因组计划不可或缺的一部分被广泛使用,已为许多克隆提供了数据库注释,为大鼠研究人员提供了丰富的BAC克隆资源,可在计算机上筛选感兴趣的基因。