1 / 5

Monomerization of Homing Endonucleases

Monomerization of Homing Endonucleases. Gene synthesis. Protein Science (2003), 12: 197-206. Library size: ~1 ×10 6. J.A.C.S. (2006), 128: 2477-84. The Length and Aa Distribution of Linkers. 19 various linkers were identified from Cre library with length from 13 Aa to 73 Aa.

Download Presentation

Monomerization of Homing Endonucleases

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. Monomerization of Homing Endonucleases Gene synthesis Protein Science (2003), 12: 197-206 Library size: ~1×106 J.A.C.S. (2006), 128: 2477-84.

  2. The Length and Aa Distribution of Linkers • 19 various linkers were identified from Cre library with length from 13 Aa to 73 Aa. • 13 various linkers were identified from Mso library with length at 13 and 33 Aa.

  3. Monomeric Mso24 • mMso24 has a 33 Aa linker between two Mso domains. • mMso24 shows comparable site-specific cleavage activity as WT Mso. • mMso24 exists as a monomer in solution, verified by size-exclusion chromatography.

  4. Thermodynamic Values of Mso24 by ITC After Correction Mso24/WTDNA WTDNA only 1. Eastburg et. al. Nucleic Acid Res. (2007), 35(21): 7209-21

  5. Current Projects • Resolve mMso24 structure. • Expression and purification of mCre3. • Crystallography study. • Biochemical study: • Binding affinity by isothermal titration calorimetry (ITC). • Thermal stability by circular dichroism (CD). • Binding and Cleavage specificity matrix. • Establish stable cell lines containing GFP-Cre and GFP-Mso cassettes. Future Projects • Engineering monomeric Cre and Mso towards novel DNA targets • using combinatorail and computational methos.

More Related