A modular, positive selection bacterial artificial chromosome vector with multiple cloning sites
A modular, positive selection bacterial artificial chromosome vector with multiple cloning sites
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DOI：
10.1006/geno.1998.5693
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发表时间：
1999-06-15
期刊：
影响因子：
4.4
通讯作者：
de Jong, PJ
中科院分区：
文献类型：
--
作者：
Frengen, E;Weichenhan, D;de Jong, PJ
为了构建用于复杂基因组测序、作图和功能研究的大型插入文库，我们构建了一种新的模块化细菌人工染色体（BAC）载体pBACe3.6（GenBank登录号U80929），该载体包含位于sacB基因内的多个克隆位点，允许在含蔗糖的培养基上对重组克隆进行正向选择。还包括 PI-SceI 核酸酶的识别位点，无论插入序列的特征如何，都可以对重组 DNA 进行线性化。 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改造 BAC 克隆。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，目前作为大规模作图和测序的资源。 (C) 1999 年学术出版社。
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. (C) 1999 Academic Press.