Open Access
Add to BibSonomyAbstract
We correct the omission of the construct and protein purification method in our recent paper [Biomed. Opt. Express 4(12), 2835–2845 (2013)].
© 2014 Optical Society of America
We add the construct and protein purification method mistakenly omitted in our recent paper [1]. The constructs were made following our previous studies [2, 3]. In brief, GST tagged Ras binding domain (RBD) and cysteine rich domain (CRD) expression constructs were generated by subcloning the two domains (amino acids 136-298) of wildtype or mutant human BRaf into pGEX4T-1 (GST-fusion vector, GE Healthcare). GST-RBD-CRD fusion proteins were induced in XL10-Gold E. coli strain (#200314, Stratagene), purified using Glutathione Sepharose 4B (#17-0756-01, GE Healthcare), and concentrated with Amicon Ultra centrifugal filters (Ultracel-30K, Millipore). Hexahistidine (His)-tagged Ras(WT) expression constructs were made by subcloning PCR-amplified full-length human H-Ras into pET28a( + ) (His-tag vector, Novagen). His-tagged Ras(WT) proteins were induced in BL21-CodonPlus (DE3)-RILP strain (#230280, Agilent Technologies), purified using Ni-NTA His-Bind Purification Kit (#70751-3, EMD Millipore Novagen), and quantified by Bradford assay kit (Bio-Rad).
References and links
1. C. Wen, A. Ye, A. Ye, and J. J. Zhu, “Single-molecule force measurement via optical tweezers reveals different kinetic features of two BRaf mutants responsible for cardio-facial-cutaneous (CFC) syndrome,” Biomed. Opt. Express 4(12), 2835–2845 (2013). [CrossRef] [PubMed]
2. Y. Qin, Y. Zhu, J. P. Baumgart, R. L. Stornetta, K. Seidenman, V. Mack, L. van Aelst, and J. J. Zhu, “State-dependent Ras signaling and AMPA receptor trafficking,” Genes Dev. 19(17), 2000–2015 (2005). [CrossRef] [PubMed]
3. A. Kielland, G. Bochorishvili, J. Corson, L. Zhang, D. L. Rosin, P. Heggelund, and J. J. Zhu, “Activity patterns govern synapse-specific AMPA receptor trafficking between deliverable and synaptic pools,” Neuron 62(1), 84–101 (2009). [CrossRef] [PubMed]