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胰岛素样生长因子(IGF)拟南芥细菌人工染色体文库的构建与鉴定

Construction and characterization of the IGF Arabidopsis BAC library.

作者信息

Mozo T, Fischer S, Shizuya H, Altmann T

机构信息

Institut für Genbiologische Forschung Berlin GmbH, Germany.

出版信息

Mol Gen Genet. 1998 Jun;258(5):562-70. doi: 10.1007/s004380050769.

Abstract

A bacterial artificial chromosome (BAC) library has been established for Arabidopsis thaliana (ecotype Col-0) covering about seven haploid nuclear genome equivalents. This library, called the Institut für Genbiologische Forschung (IGF) BAC library, consists of 10,752 recombinant clones carrying inserts (generated by partial EcoRI digestion) of an average size of about 100 kb in a modified BAC vector, pBeloBAC-Kan. Hybridization with organellar DNA and nuclear repetitive DNA elements revealed the presence of 1.1% clones with mitochondrial DNA, 0.2% clones with plastid DNA, 3.2% clones with the 180 bp paracentromeric repeat, 1.6% clones with 5S rDNA, and 10.8% clones with the 18S-25S rDNA repeat. With its extensive genome coverage, its rather uniformly sized inserts (80 kb < 85% < 120 kb) and low contamination with organellar DNA, this library provides an excellent resource for A. thaliana genomic mapping, map-based gene cloning, and genome sequencing.

摘要

已构建了一个拟南芥(生态型Col-0)的细菌人工染色体(BAC)文库,其覆盖约7个单倍体核基因组当量。该文库称为德国分子生物学研究所(IGF)BAC文库,由10752个重组克隆组成,这些克隆在改良的BAC载体pBeloBAC-Kan中携带平均大小约为100 kb的插入片段(通过EcoRI部分酶切产生)。与细胞器DNA和核重复DNA元件的杂交显示,有1.1%的克隆含有线粒体DNA,0.2%的克隆含有质体DNA,3.2%的克隆含有180 bp的近着丝粒重复序列,1.6%的克隆含有5S rDNA,10.8%的克隆含有18S-25S rDNA重复序列。该文库具有广泛的基因组覆盖范围、相当均匀大小的插入片段(80 kb<85%<120 kb)以及低细胞器DNA污染率,为拟南芥基因组作图、基于图谱的基因克隆和基因组测序提供了极好的资源。

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