Tandem affinity purification vectors for use in gram positive bacteria.

Yang X; Doherty GP; Lewis PJ

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Tandem affinity purification vectors for use in gram positive bacteria.

机译：用于革兰氏阳性细菌的串联亲和纯化载体。

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Tandem affinity purification has become a valuable tool for the isolation of protein complexes. Here we describe the construction and use of a series of plasmid vectors for Gram positive bacteria. The vectors utilize the SPA tag as well as variants containing a 3C rather than the TEV protease site as 3C protease has been shown to work efficiently at the low temperatures (4 degrees C) used to isolate protein complexes. In addition, a further vector incorporates a GST moiety in place of the 3xFLAG of the SPA tag which provides an additional tagging option for situations where SPA binding may be inefficient. The vectors are all compatible with previously constructed fluorescent protein fusion vectors enabling construction of a suite of affinity and fluorescently tagged genes using a single PCR product.

机译：串联亲和纯化已成为分离蛋白质复合物的重要工具。在这里，我们描述了革兰氏阳性细菌的一系列质粒载体的构建和使用。载体利用SPA标签以及含有3C而不是TEV蛋白酶位点的变体，因为已显示3C蛋白酶在用于分离蛋白质复合物的低温（4℃）下有效地起作用。此外，另一种载体掺入了GST部分来代替SPA标签的3xFLAG，这为SPA结合效率低下的情况提供了额外的标签选择。这些载体均与先前构建的荧光蛋白融合载体兼容，从而能够使用单个PCR产物构建一组亲和力和荧光标记的基因。

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《Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems》 |2008年第1期|共9页
作者
Yang X; Doherty GP; Lewis PJ;
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正文语种 eng
中图分类医学遗传学;
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1. Tandem affinity purification vectors for use in gram positive bacteria. [J] . Yang X, Doherty GP, Lewis PJ Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems . 2008,第1期

机译：用于革兰氏阳性细菌的串联亲和纯化载体。
2. Isolation and purification of enterocin E-760 with broad antimicrobial activity against gram-positive and gram-negative bacteria. [J] . Line JE, Svetoch EA, Eruslanov BV, Antimicrobial agents and chemotherapy. . 2008,第3期

机译：肠毒素E-760的分离和纯化对革兰氏阳性和革兰氏阴性细菌具有广泛的抗菌活性。
3. Improved vectors for nisin-controlled expression in gram-positive bacteria. [J] . Bryan EM, Bae T, Kleerebezem M, Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems . 2000,第2期

机译：用于革兰氏阳性细菌中乳链菌肽控制表达的改良载体。
4. Purification and Characterization of the Human COP9 Signalosome Using a New Tandem Affinity Purification Strategy and Quantitative Mass Spectrometry [C] . Lei Fang, Xiaorong Wang, Phang-lang Chen, ASMS Conference on Mass Spectrometry and Allied Topics . 2008

机译：使用新的串联亲和纯化策略和定量质谱法纯化和表征人COP9信号组
5. Iron uptake in Gram-negative and Gram-positive bacteria. [D] . Jin, Bo. 2006

机译：革兰氏阴性和革兰氏阳性细菌中的铁吸收。
6. A pair of mobilizable shuttle vectors conferring resistance to spectinomycin for molecular cloning in Escherichia coli and in gram-positive bacteria. [O] . P Trieu-Cuot, C Carlier, C Poyart-Salmeron, 1990

机译：一对赋予大观霉素抗性的可动穿梭载体用于在大肠杆菌和革兰氏阳性细菌中进行分子克隆。
7. A pair of mobilizable shuttle vectors conferring resistance to spectinomycin for molecular cloning in Escherichia coli and in gram-positive bacteria. [O] . Trieu-Cuot, P, Carlier, C, Poyart-Salmeron, C, 1990

机译：一对赋予大观霉素抗性的可动穿梭载体，用于在大肠杆菌和革兰氏阳性细菌中进行分子克隆。

Tandem affinity purification vectors for use in gram positive bacteria.

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