Improvement of bacterial two-hybrid vectors for detection of fusion proteins and transfer to pBAD-tandem affinity purification, calmodulin binding peptide, or 6-histidine tag vectors
Improvement of bacterial two-hybrid vectors for detection of fusion proteins and transfer to pBAD-tandem affinity purification, calmodulin binding peptide, or 6-histidine tag vectors
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DOI：
10.1002/pmic.200800270
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发表时间：
2008-11-01
期刊：
影响因子：
3.4
通讯作者：
Bouveret, Emmanuelle
中科院分区：
文献类型：
--
作者：
Battesti, Aurelia;Bouveret, Emmanuelle
Karimova等人在1998年开发的细菌双杂交技术的原始载体不能检测重组蛋白。在这里，我们提出了两种方法来解决这个问题，要么使用含有Flag表位的新质粒，要么使用一种技巧来检测腺苷酸环化酶的T18结构域。此外，我们还描述了一组与我们的双杂交载体具有相同克隆位点的TAP， CBP或6-组氨酸标记载体。
The original vectors of the bacterial two-hybrid technique developed by Karimova et al. in 1998 did not enable detection of the recombinant proteins. Here, we propose two methods resolving this problem, either using new plasmids containing the Flag epitope, or using a trick to detect the T18 domain of adenylate cyclase. Furthermore, we describe a set of vectors for TAP, CBP or 6-histidine tagging that possess the same cloning site as our two-hybrid vectors.