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人类2号染色体显微切割克隆衍生的序列标签位点的产生与定位

Generation and mapping of human chromosome 2 microdissection clone-derived STSs.

作者信息

Gingrich J C, Garnes J A, Lee D A, Boehrer D M

机构信息

Human Genome Center, Lawrence Livermore National Laboratory, Livermore, CA, USA.

出版信息

Cytogenet Cell Genet. 1996;74(4):272-6. doi: 10.1159/000134433.

Abstract

Twenty-nine new sequence-tagged sites (STSs) were derived from DNA sequences of clones from two human chromosome 2 microdissection libraries. The specificity of the STSs for human chromosome 2 was first demonstrated by PCR amplification of DNA from genomic human and hamster cells and a human chromosome 2-containing human x hamster hybrid cell line. The STSs were then mapped to chromosome 2 by two different approaches. In the first attempt, 12 of the STSs were shown to PCR amplify YAC clones associated with genetic markers on the chromosome. In the second approach, 27 of the STSs were localized to chromosome bands by FISH using cosmid or PAC clones encoding the STSs. The specific STSs mapped to chromosome 2 by these two approaches tie together the genetic and cytogenetic maps of the chromosome at the two termini. The distribution of these STSs further defines the region of the chromosome present in the two microdissection libraries.

摘要

29个新的序列标签位点(STS)源自两个人类2号染色体显微切割文库中克隆的DNA序列。通过对人类基因组DNA、仓鼠细胞DNA以及一个含有2号染色体的人-仓鼠杂种细胞系进行PCR扩增,首次证明了这些STS对人类2号染色体的特异性。然后通过两种不同的方法将这些STS定位到2号染色体上。在首次尝试中,12个STS能通过PCR扩增与该染色体上遗传标记相关的酵母人工染色体(YAC)克隆。在第二种方法中,使用编码这些STS的黏粒或噬菌体P1人工染色体(PAC)克隆,通过荧光原位杂交(FISH)将27个STS定位到染色体带。通过这两种方法定位到2号染色体上的特定STS将该染色体两端的遗传图谱和细胞遗传图谱联系在一起。这些STS的分布进一步界定了两个显微切割文库中所包含的染色体区域。

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