Talk:CH391L/S12/Fluorescent Proteins: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
No edit summary
Line 1: Line 1:
*'''[[User:Peter Otoupal|Peter Otoupal]] 19:05, 25 March 2012 (EDT)''':Are the GFP variants now used in laboratories typically Enhanced GFP, or are the "original" classes still mainly used? Also, how did Zhang et al know to target the Ser65 and Phe64 to improve fluorescence? Or did they just create a library and cross their fingers? Thanks!
*'''[[User:Peter Otoupal|Peter Otoupal]] 19:05, 25 March 2012 (EDT)''':Are the GFP variants now used in laboratories typically Enhanced GFP, or are the "original" classes still mainly used? Also, how did Zhang et al know to target the Ser65 and Phe64 to improve fluorescence? Or did they just create a library and cross their fingers? Thanks!
**'''[[User:Midhat Patel|Midhat Patel]] 03:56, 26 March 2012 (EDT)''': The variants used are primarily GFP at this point. The amino acids in locations 64-67 were targeted because random mutation libraries showed that when these were altered, fluorescence either disappeared or was different, so these were the critical amino acids for fluorescence.


*'''[[User:Joe Hanson|Joe Hanson]] 16:50, 19 March 2012 (EDT)''': Can you add something about Split GFP? It's a pretty cool system for things like solubility evolution, and it allows you to use smaller fusion proteins when compared to attaching the entire GFP ORF.
*'''[[User:Joe Hanson|Joe Hanson]] 16:50, 19 March 2012 (EDT)''': Can you add something about Split GFP? It's a pretty cool system for things like solubility evolution, and it allows you to use smaller fusion proteins when compared to attaching the entire GFP ORF.

Revision as of 00:56, 26 March 2012

  • Peter Otoupal 19:05, 25 March 2012 (EDT):Are the GFP variants now used in laboratories typically Enhanced GFP, or are the "original" classes still mainly used? Also, how did Zhang et al know to target the Ser65 and Phe64 to improve fluorescence? Or did they just create a library and cross their fingers? Thanks!
    • Midhat Patel 03:56, 26 March 2012 (EDT): The variants used are primarily GFP at this point. The amino acids in locations 64-67 were targeted because random mutation libraries showed that when these were altered, fluorescence either disappeared or was different, so these were the critical amino acids for fluorescence.
  • Joe Hanson 16:50, 19 March 2012 (EDT): Can you add something about Split GFP? It's a pretty cool system for things like solubility evolution, and it allows you to use smaller fusion proteins when compared to attaching the entire GFP ORF.
    • Midhat Patel 03:30, 26 March 2012 (EDT): A section has been added about Split GFP.
  • Jeffrey E. Barrick 13:15, 25 March 2012 (EDT): Is there a destabilized GFP that works in bacteria in the iGEM registry? How does it work?
  • Jeffrey E. Barrick 13:20, 25 March 2012 (EDT): This statement under EGFP is confusing "the altered enzymes in EGFP". I believe they are talking about the expression of an enzyme that has been fused to EGFP in this case, since a common application of GFP is to fuse it to a protein to examine localization of that protein in the cell.
    • Midhat Patel 03:36, 26 March 2012 (EDT): I apologize, that's an embarrassing typo. What I meant to say was that the altered amino acids are those that are more common in eukaryotic cells, I have corrected the statement.
  • Jeffrey E. Barrick 13:32, 25 March 2012 (EDT): Page could be improved by making the red links go to articles on Wikipedia.
    • Midhat Patel 03:35, 26 March 2012 (EDT): Taken care of, I didn't realize it was linking to non-existent Open WetWare pages and not the Wiki pages themselves.