Frengen E, Weichenhan D, Zhao B, Osoegawa K, van Geel M, de Jong P J
Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
Genomics. 1999 Jun 15;58(3):250-3. doi: 10.1006/geno.1998.5693.
To construct large-insert libraries for the sequencing, mapping, and functional studies of complex genomes, we have constructed a new modular bacterial artificial chromosome (BAC) vector, pBACe3.6 (GenBank Accession No. U80929). This vector contains multiple cloning sites located within the sacB gene, allowing positive selection for recombinant clones on sucrose-containing medium. A recognition site for the PI-SceI nuclease has also been included, which permits linearization of recombinant DNA irrespective of the characteristics of the insert sequences. An attTn7 sequence present in pBACe3.6 permits retrofitting of BAC clones by Tn7-mediated insertion of desirable sequence elements into the vector portion. The ability to retrofit BAC clones will be useful for functional analysis of genes carried on the cloned inserts. The pBACe3.6 vector has been used for the construction of many genomic libraries currently serving as resources for large-scale mapping and sequencing.
为构建用于复杂基因组测序、图谱绘制和功能研究的大插入片段文库，我们构建了一种新型模块化细菌人工染色体（BAC）载体pBACe3.6（GenBank登录号U80929）。该载体在sacB基因内含有多个克隆位点，可在含蔗糖培养基上对重组克隆进行阳性筛选。还包含PI-SceI核酸酶的识别位点，这使得重组DNA能够线性化，而与插入序列的特性无关。pBACe3.6中存在的attTn7序列允许通过Tn7介导将所需序列元件插入载体部分来对BAC克隆进行改造。改造BAC克隆的能力将有助于对克隆插入片段上携带的基因进行功能分析。pBACe3.6载体已用于构建许多基因组文库，目前这些文库作为大规模图谱绘制和测序的资源。
