Yu J, Tong S, Whittier A, Kao F T
Eleanor Roosevelt Institute for Cancer Research, Denver, Colorado 80206, USA.
Somat Cell Mol Genet. 1995 Sep;21(5):335-43. doi: 10.1007/BF02257468.
Three region-specific libraries have been constructed from the long arm of human chromosome 2, including regions 2q33-35 (2Q2 library), 2q31-32 (2Q3) and 2q23-24 (2Q4). Chromosome microdissection and the MboI linker-adaptor microcloning techniques were used in constructing these libraries. The libraries comprised hundreds of thousands of microclones in each library. Approximately half of the microclones in the library contained unique or low-copy number sequence inserts. The insert sizes ranged between 50 and 800 bp, with a mean of 130-190 bp. Southern blot analysis of individual unique sequence microclones showed that 70-94% of the microclones were derived from the dissected region. 31 unique sequence microclones from the 2Q2 library, 31 from 2Q3, and 30 from 2Q4, were analyzed for insert sizes, the hybridizing genomic HindIII fragment sizes, and cross-hybridization to rodent species. These libraries and the short insert microclones derived from the libraries should be useful for high resolution physical mapping, sequence-ready reagents for large scale genomic sequencing, and positional cloning of disease-related genes assigned to these regions, e.g. the recessive familial amyotrophic lateral sclerosis assigned to 2q33-q35, and a type I diabetes susceptibility gene to 2q31-q33.
已从人类2号染色体长臂构建了三个区域特异性文库，包括2q33 - 35区域（2Q2文库）、2q31 - 32区域（2Q3文库）和2q23 - 24区域（2Q4文库）。构建这些文库时使用了染色体显微切割和MboI接头 - 衔接子微克隆技术。每个文库包含数十万微克隆。文库中大约一半的微克隆含有独特或低拷贝数序列插入片段。插入片段大小在50至800 bp之间，平均为130 - 190 bp。对单个独特序列微克隆的Southern印迹分析表明，70 - 94%的微克隆源自切割区域。对2Q2文库的31个独特序列微克隆、2Q3文库的31个以及2Q4文库的30个微克隆进行了插入片段大小、杂交基因组HindIII片段大小以及与啮齿动物物种交叉杂交的分析。这些文库以及从文库衍生的短插入微克隆应有助于高分辨率物理图谱绘制、用于大规模基因组测序的序列就绪试剂以及定位克隆定位于这些区域的疾病相关基因，例如定位于2q33 - q35的隐性家族性肌萎缩侧索硬化症以及定位于2q31 - q33的I型糖尿病易感基因。
