Construction of compatible wide-host-range shuttle vectors for lactic acid bacteria and Escherichia coli
Construction of compatible wide-host-range shuttle vectors for lactic acid bacteria and Escherichia coli
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DOI：
10.1006/plas.2001.1531
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发表时间：
2001-09-01
期刊：
影响因子：
2.6
通讯作者：
Pérez-Martínez, G
中科院分区：
文献类型：
--
作者：
Pérez-Arellano, I;Zúñiga, M;Pérez-Martínez, G
构建了新的穿梭克隆载体集合，可用于广泛的宿主范围，因为它们携带在大肠杆菌中具有功能的复制起点（p15A、pWV01、ColE1）。乳酸乳球菌、乳酸杆菌和枯草芽孢杆菌（pAM beta1、pWV01）。这些质粒包含来自 pJDC9 的 lacZ-T1T2 盒，它允许 X-gal 选择和克隆可能导致大肠杆菌中质粒不稳定的 DNA 片段。此外，它们已被证明在干酪乳杆菌中具有结构和分离稳定性，其中它们的拷贝数已通过实时定量 PCR 测定。此外，抗生素抗性标记（fl-内酰胺酶、氯霉素乙酰转移酶和红霉素转乙酰酶）以及theta和滚环复制起点已被组合以获得这组可在乳酸菌和大肠杆菌中共转化的相容质粒（pIA家族）。 (C) 2001 年学术出版社。
A new collection of shuttle cloning vectors has been constructed that can be used in a broad host range, because they carry replication origins which are functional in Escherichia coli (p15A, pWV01, ColE1). Lactococcus lactis, lactobacilli, and Bacillus subtilis (pAM beta1, pWV01). These plasmids, contain the lacZ-T1T2 cassette from pJDC9, which allows the X-gal selection and cloning of DNA fragments that could cause plasmid instability in E. coli. In addition, they have been proved to be structurally and segregationally stable in Lactobacillus casei, in which their copy number has been determined by real-time quantitative PCR. Furthermore, the antibiotic resistance markers (fl-lactamase, chloramphenicol acetyl transferase, and erythromycin transacetylase) and the theta and rolling circle replicating origins have been combined to obtain this set of compatible plasmids (pIA family) that can be cotransformed, both in lactic acid bacteria and in E. coli. (C) 2001 Academic Press.