Chromoprotein Mutagenesis

This has not been tried yet. I have been thinking of a way include generating mutations in the lab.

The general idea I have in mind is using PCR mutagenesis (error prone PCR) to make haphazard changes to chromoproteins, clone these into E. coli, select clones with interesting novel phenotypes, then sequence the insert to find the mutation responsible.

Literature

Liljeruhm, J., Funk, S. K., Tietscher, S., Edlund, A. D., Jamal, S., Wistrand-Yuen, P., ... & Törnblom, V. (2018). Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology. Journal of Biological Engineering, 12(1), 8; https://jbioleng.biomedcentral.com/articles/10.1186/s13036-018-0100-0 ; https://scholar.google.com/scholar?cluster=17927173279619010872

McCullum, E. O., Williams, B. A., Zhang, J., & Chaput, J. C. (2010). Random mutagenesis by error-prone PCR. In In Vitro Mutagenesis Protocols (pp. 103-109). Humana Press, Totowa, NJ; https://www.researchgate.net/profile/Sibtain_Afzal/post/Is_anyone_familiar_with_random_mutagenesis2/attachment/59d62a00c49f478072e9cbe3/AS%3A272472463609862%401441973905587/download/Random+Mutagenesis+by+Error-Prone+PCR.pdf ; https://scholar.google.com/scholar?cluster=14328399351368807250

Roberts, T. M., Rudolf, F., Meyer, A., Pellaux, R., Whitehead, E., Panke, S., & Held, M. (2016). Identification and characterisation of a pH-stable GFP. Scientific Reports, 6, 28166; https://www.nature.com/articles/srep28166 ; https://scholar.google.com/scholar?cluster=13704240519438793468

Ruller, R., Silva‐Rocha, R., Silva, A., Cruz Schneider, M. P., & Ward, R. J. (2011). A practical teaching course in directed protein evolution using the green fluorescent protein as a model. Biochemistry and Molecular Biology Education, 39(1), 21-27; https://iubmb.onlinelibrary.wiley.com/doi/full/10.1002/bmb.20430 ; https://scholar.google.com/scholar?cluster=10821652129227038353

Addgene. Protocols: Plasmid Cloning by PCR. Accessed July 18, 2018, https://www.addgene.org/protocols/pcr-cloning/