一个用于对完整人类基因组进行测序的细菌人工染色体文库。
A bacterial artificial chromosome library for sequencing the complete human genome.
作者信息
Osoegawa K, Mammoser A G, Wu C, Frengen E, Zeng C, Catanese J J, de Jong P J
机构信息
Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
出版信息
Genome Res. 2001 Mar;11(3):483-96. doi: 10.1101/gr.169601.
A 30-fold redundant human bacterial artificial chromosome (BAC) library with a large average insert size (178 kb) has been constructed to provide the intermediate substrate for the international genome sequencing effort. The DNA was obtained from a single anonymous volunteer, whose identity was protected through a double-blind donor selection protocol. DNA fragments were generated by partial digestion with EcoRI (library segments 1--4: 24-fold) and MboI (segment 5: sixfold) and cloned into the pBACe3.6 and pTARBAC1 vectors, respectively. The quality of the library was assessed by extensive analysis of 169 clones for rearrangements and artifacts. Eighteen BACs (11%) revealed minor insert rearrangements, and none was chimeric. This BAC library, designated as "RPCI-11," has been used widely as the central resource for insert-end sequencing, clone fingerprinting, high-throughput sequence analysis and as a source of mapped clones for diagnostic and functional studies.
已构建了一个具有30倍冗余且平均插入片段大小较大（178 kb）的人类细菌人工染色体（BAC）文库，为国际基因组测序工作提供中间底物。DNA取自一名匿名志愿者，通过双盲供体选择方案保护其身份。用EcoRI（文库片段1 - 4：24倍）和MboI（片段5：6倍）进行部分消化产生DNA片段，并分别克隆到pBACe3.6和pTARBAC1载体中。通过对169个克隆进行重排和人工产物的广泛分析来评估文库质量。18个BAC（11%）显示出轻微的插入重排，没有一个是嵌合的。这个被命名为“RPCI - 11”的BAC文库已被广泛用作插入末端测序、克隆指纹分析、高通量序列分析的核心资源，以及用于诊断和功能研究的定位克隆来源。
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