Lissa1: Project Page

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Contents

What is the Point of this Project?

Debugging!

The goal of this project is to characterize the inner workings of the phospholocator , designed and built by Samantha Sutton. Although we know how the phospholocator device works, we don't have a quantitative description of how the parts interact.

Since the effectiveness (gain) of the phospholocator (PPL) device depends on the kinase parts being active, or phosphorylated, I will spend this summer finding out:

  • how much phosphorylated kinase is present in a cell
  • the relationship between phosphorylated kinase and phosphorylated PPL
  • how changing the concentration of active kinase and PPL in the cell effects the gain of the system
  • how to use this knowledge (which will be collected and posted on the [Registry of Standard Biological Parts]) to increase the gain of the PPL

How To...

  1. How to Monitor Cdc28 and Fus3 levels in yeast: Image:Fus3cdc28.doc
  2. How to Get Linear Expression of Phospholocator: Image:Linearexpr.doc

Protocols:


Plan of Attack

Current Thinking

Schedule for June 14 - June 23

Schedule for June 24 - June 30

Schedule for July 1 - July 8

Schedule for July 9 - July 16

Schedule for July 17 - July 26

Schedule for July 27 - August 2

Schedule for August 6 - August 14

References

Swi5 NLS

  1. Moll T, Tebb G, Surana U, Robitsch H, and Nasmyth K. . pmid:1652372. PubMed HubMed [Swi5]


Fus3 Docking Site

  1. Bardwell AJ, Flatauer LJ, Matsukuma K, Thorner J, and Bardwell L. . pmid:11134045. PubMed HubMed [Fus3Dock]

Swapping Docking Sites

  1. Smith JA, Poteet-Smith CE, Lannigan DA, Freed TA, Zoltoski AJ, and Sturgill TW. . pmid:10922375. PubMed HubMed [Swap]

Oriented Peptide Library Screens

  1. Obata T, Yaffe MB, Leparc GG, Piro ET, Maegawa H, Kashiwagi A, Kikkawa R, and Cantley LC. . pmid:10945990. PubMed HubMed [YaffeCantley]

Chemical Inhibitors of Kinase function

  1. Bishop AC, Ubersax JA, Petsch DT, Matheos DP, Gray NS, Blethrow J, Shimizu E, Tsien JZ, Schultz PG, Rose MD, Wood JL, Morgan DO, and Shokat KM. . pmid:11014197. PubMed HubMed [Shokat]

Gel Shift Paper

  1. Li E, Meldrum E, Stratton HF, and Stone DE. . pmid:9539416. PubMed HubMed [Shift]

Cool Modular Protein Engineering Examples

Error fetching PMID 16602891:
  1. Error fetching PMID 16602891: [Overview]
  2. Dueber JE, Yeh BJ, Chak K, and Lim WA. . pmid:14512628. PubMed HubMed [Allostery]
  3. Park SH, Zarrinpar A, and Lim WA. . pmid:12511654. PubMed HubMed [Scaffold]
All Medline abstracts: PubMed HubMed
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