Birren B W, Tachi-iri Y, Kim U J, Nguyen M, Shizuya H, Korenberg J R, Simon M I
Division of Biology 147-75, California Institute of Technology, Pasadena, California, 91125, USA.
Genomics. 1996 May 15;34(1):97-106. doi: 10.1006/geno.1996.0246.
We have created a resource for chromosome 22 consisting of 96 unique, well-characterized Fosmids. The Fosmid vector permits efficient cloning of DNA fragments averaging 40 kb in a single-copy vector based on the F factor of Escherichia coli. We have found that Fosmid clones from human chromosome 22 show remarkable stability and are useful for a wide variety of applications in genome analysis. These 96 clones have been localized by FISH, using high-resolution fluorescent banding and multicolor mapping techniques, and their position on the chromosome was correlated with their content of a number of common repeated sequence elements. We identified a subset of clones likely to contain genes by restriction analysis using the enzymes NotI, MluI, SacII, and BssHII. This collection of cytogenetically anchored clones, representing nearly 7% of the chromosome, is of immediate value for detecting chromosomal rearrangements, for use in gene isolation, and as a framework for physical mapping.
我们创建了一个22号染色体资源库，其中包含96个独特且特征明确的Fosmid克隆。Fosmid载体能够基于大肠杆菌的F因子，在单拷贝载体中高效克隆平均长度为40 kb的DNA片段。我们发现，来自人类22号染色体的Fosmid克隆表现出显著的稳定性，可用于基因组分析中的多种应用。利用高分辨率荧光带型和多色图谱技术，通过荧光原位杂交（FISH）对这96个克隆进行了定位，并将它们在染色体上的位置与其所含多种常见重复序列元件的情况相关联。我们使用NotI、MluI、SacII和BssHII酶通过限制性分析鉴定出了一个可能包含基因的克隆子集。这组细胞遗传学定位的克隆代表了该染色体近7%的区域，对于检测染色体重排、用于基因分离以及作为物理图谱构建的框架具有直接价值。
