Sodium chloride 'AnalaR'

公司名称: VWR

产品编号: 102415K

Protocol for the High-quality Plasmid Isolation from Different Recalcitrant Bacterial Species: Agrobacterium spp., Rhizobium sp., and Bacillus thuringiensis

Preshobha Kodackattumannil, Shina Sasi, Saranya Krishnan, Geetha Lekshmi, Martin Kottackal and Khaled M. A. Amiri

Published online: 2023-08-05

Bio-protocol()

Company-protocol()

Other protocol()

Evaluation of Plasmid Stability by Negative Selection in Gram-negative Bacteria

Author:

Damián Lobato Márquez and Laura Molina García,

Date:

2017-05-05

[Abstract] Plasmid stability can be measured using antibiotic-resistance plasmid derivatives by positive selection. However, highly stable plasmids are below the sensitivity range of these assays. To solve this problem we describe a novel, highly sensitive method to measure plasmid stability based on the selection of plasmid-free cells following elimination of plasmid-containing cells. The assay proposed here is based on an aph-parE cassette. When synthesized in the cell, the ParE toxin induces cell death. ParE synthesis is controlled by a rhamnose-inducible promoter. When bacteria carrying the aph-parE ... [摘要] 可以通过阳性选择使用抗生素抗性质粒衍生物来测量质粒稳定性。然而，高度稳定的质粒低于这些测定的灵敏度范围。为了解决这个问题，我们描述了一种新颖的，高度灵敏的方法来测量质粒稳定性，这是基于在含有质粒的细胞消除后，无质粒细胞的选择。这里提出的检测方法是基于aph-parE 盒式磁带。当细胞合成时，ParE毒素诱导细胞死亡。 ParE合成由鼠李糖诱导型启动子控制。当携带 aph-parE 模块的细菌生长在含有鼠李糖作为唯一碳源的培养基中时，合成ParE，消除含有质粒的细胞。进一步使用卡那霉素抗性（ aph ）来证实在鼠李糖生长的细菌中不存在质粒。

背景通过使用抗生素抗性质粒衍生物的阳性选择测定质粒稳定性。携带研究质粒的细胞在选择性抗生素（Gerdes等人，1985; del Solar等人，1987）的存在下被阳性选择。这种技术的主要缺点是其灵敏度;高度稳定的质粒低于这些测定的敏感性。为了解决这个问题，已经描述了依靠直接选择无质粒细胞例如 - 四环素系统的替代方法（Bochner等人，1980; Maloy和Nunn，1981; Garcia-Quintanilla 等人，2006）。 - ...

产品评论

Sodium chloride 'AnalaR'

免责声明