Lange Cornelia, Holtgräwe Daniela, Schulz Britta, Weisshaar Bernd, Himmelbauer Heinz
Max Planck Institute for Molecular Genetics, Ihnestr. 63-73, 14195 Berlin, Germany.
Genome. 2008 Nov;51(11):948-51. doi: 10.1139/G08-071.
A sugar beet (Beta vulgaris) fosmid library from the doubled haploid accession KWS2320 encompassing 115 200 independent clones was constructed and characterized. The average insert size of the fosmid library was determined by pulsed field gel electrophoresis to be 39 kbp on average, thus representing 5.9-fold coverage of the sugar beet genome (758 Mbp). PCR screening of plate pools with primer pairs against nine sugar beet genes supported the insert size estimation. BLAST searches with 2951 fosmid end-sequences originating from 1510 clones (1536 clones attempted) revealed little contamination with organellar DNA (2.1% chloroplast DNA, 0.3% mitochondrial DNA). The sugar beet fosmid library will be integrated in the presently ongoing efforts to determine the sequence of the sugar beet genome. Fosmids will be publicly available in the format of plate pools and individual clones.
构建并表征了一个来自双单倍体系KWS2320的甜菜（Beta vulgaris）fosmid文库，该文库包含115200个独立克隆。通过脉冲场凝胶电泳确定该fosmid文库的平均插入片段大小平均为39 kbp，因此代表了甜菜基因组（758 Mbp）5.9倍的覆盖率。用针对九个甜菜基因的引物对进行平板池PCR筛选支持了插入片段大小的估计。对来自1510个克隆（尝试了1536个克隆）的2951个fosmid末端序列进行BLAST搜索，结果显示细胞器DNA污染很少（叶绿体DNA占2.1%，线粒体DNA占0.3%）。该甜菜fosmid文库将整合到目前正在进行的甜菜基因组测序工作中。Fosmids将以平板池和单个克隆的形式公开提供。
