Wang M, Chen X N, Shouse S, Manson J, Wu Q, Li R, Wrestler J, Noya D, Sun Z G, Korenberg J
Department of Pharmacology, University of North Carolina at Chapel Hill 27599-7365.
Genomics. 1994 Dec;24(3):527-34. doi: 10.1006/geno.1994.1662.
We have constructed a human chromosome 2-specific bacterial artificial chromosome (BAC) library using DNA from the somatic cell hybrid GM10826. The average size of the clones is about 63 kb. The coverage and distribution of the library were estimated by screening with known polymorphic genetic markers and fluorescence in situ hybridization (FISH). Twenty-one markers tested positive when DNA pools prepared from approximately one-sixth of the library were screened with 33 known markers. This is consistent with the theoretical calculation of 63% coverage at one genomic equivalent. This suggested that the coverage of the library is approximately 5-6x. FISH analysis with 54 BACs revealed single site hybridization to chromosome 2, and the clones were distributed randomly on the chromosome. We have also performed direct sequencing of the BAC insert ends to generate sequence-tagged sites suitable for mapping and chromosome walking. This is the first reported human chromosome 2-specific BAC library and should provide a resource for physical mapping and disease searching for this chromosome.
我们利用体细胞杂种GM10826的DNA构建了一个人类2号染色体特异性细菌人工染色体（BAC）文库。克隆的平均大小约为63 kb。通过用已知的多态性遗传标记进行筛选以及荧光原位杂交（FISH）来估计文库的覆盖率和分布。当用33个已知标记对从约六分之一的文库制备的DNA池进行筛选时，21个标记呈阳性。这与在一个基因组当量下63%覆盖率的理论计算结果一致。这表明该文库的覆盖率约为5 - 6倍。对54个BAC进行的FISH分析显示在2号染色体上有单一位点杂交，并且这些克隆随机分布在染色体上。我们还对BAC插入片段末端进行了直接测序，以生成适用于作图和染色体步移的序列标签位点。这是首个报道的人类2号染色体特异性BAC文库，应为该染色体的物理作图和疾病研究提供资源。
