Part:BBa_K4020007

Usage and Biology

LexA Binding Domain (pBT3-N) SID (TEV site linker included) is a DNA sequence that encodes SID linker from SALSA protein, Tobacco Etch Virus (TEV) protease recognition site (Nam et al., 2020), followed by LexA Binding Domain. The latter is from the LexA protein, a transcriptional repressor (Zhang et al., 2010) of the SOS regulon in Escherichia coli (Thliveris & Mount, 1992) that allows the binding of different regulatory molecules. SIDs are roughly 20-amino-acid-long threonine-serine-proline-rich stretches separating the SRCR domains in the SALSA protein (Reichhardt et al., 2020; Turenchalk & Xu, 2001). TEV linker codes for the sequence of amino acids ENLYFQG/S, which TEV protease recognizes and cleaves between Q and G/S (Nam et al., 2020). It was acquired from the DualMembrane Kit 3 (Thaminy et al., 2003).

References

• Nam, H., Hwang, B. J., Choi, D., Shin, S., & Choi, M. (2020). Tobacco etch virus (TEV) protease with multiple mutations to improve solubility and reduce self‐cleavage exhibits enhanced enzymatic activity. FEBS Open Bio, 10(4), 619. https://doi.org/10.1002/2211-5463.12828
• Reichhardt, M. P., Loimaranta, V., Lea, S. M., & Johnson, S. (2020). Structures of SALSA/DMBT1 SRCR domains reveal the conserved ligand-binding mechanism of the ancient SRCR fold. Life Science Alliance, 3(4). https://doi.org/10.26508/LSA.201900502
• Thliveris, A. T., & Mount, D. W. (1992). Genetic identification of the DNA binding domain of Escherichia coli LexA protein. Proceedings of the National Academy of Sciences, 89(10), 4500–4504. https://doi.org/10.1073/PNAS.89.10.4500
• Turenchalk, G. S., & Xu, T. (2001). Lats in Cell-cycle Regulation and Tumorigenesis BT - Encyclopedic Reference of Cancer. Encyclopedic Reference of Cancer, 491–496. https://doi.org/10.1007/3-540-30683-8_944
• Zhang, A. P. P., Pigli, Y. Z., & Rice, P. A. (2010). Structure of the LexA-DNA complex and implications for SOS box measurement. Nature, 466(7308), 883. https://doi.org/10.1038/NATURE09200
• Thaminy, S., Auerbach, D., Arnoldo, A., & Stagljar, I. (2003). Identification of Novel ErbB3-Interacting Factors Using the  Split-Ubiquitin Membrane Yeast Two-Hybrid System. Genome Research, 13(7), 1744. https://doi.org/10.1101/GR.1276503