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  2. TOX-R Dimerization Assay

TOX-R Dimerization AssayTOX-R 二聚化测定

释义:

TOX-R Dimerization Assay is used to identify the strength of interactions in the transmembrane domains within the E. coli membrane. The assay is achieved by constructing a chimeric ToxR-TM-MBP (composed of ToxR's DNA binding/transcriptional activation domain, the transmembrane domain of interest (TM), and maltose-binding protein). The binding of two TM domains activates ToxR-mediated reporter gene expression (e.g., CAT, chloramphenicol acetyltransferase activity), and the level can be quantitatively analyzed by measuring CAM resistance or CAT activity in cell-free extracts.

TOX-R 二聚化测定可用于鉴定大肠杆菌 (E. coli) 内膜中跨膜域的相互作用强度。通过嵌合构建体 ToxR-TM-MBP (由 ToxR 的 DNA 结合/转录激活域、感兴趣的跨膜域 (TM) 和麦芽糖结合蛋白组成) 实现。两个 TM 的结合可激活 ToxR 介导的报告基因 (如 CAT,氯霉素乙酰基转移酶活性) 的表达,其水平可以通过测定 CAM 抗性或无细胞提取物中的 CAT 活性来定量分析。

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