BE.109:Bio-material engineering

Module 4

Instructors: Angela Belcher and Natalie Kuldell

TA: Eileen Higham

In this experimental module you will study an unusual protein, one that allows yeast to bind a metal, gold. Over the next few weeks you will purify yeast based on this binding property, and then you’ll vary some experimental condition to improve the yeast/gold interaction. Using your optimized conditions, you will screen a library of yeast to isolate a new gold-binding strain. The DNA encoding your new gold-binding protein will be sequenced and, with any luck, you’ll elucidate some amino acid requirements for the yeast/metal interaction.

Lab handouts

Day 1: Screening library

Day 2: Optimizing panning

Day 3: Rescreening gold binders

Day 4: PCR of gold binding candidates

Day 5: Student presentations

Day 6: Sequence analysis

References

Note: PDF reprints are provided below within the context of fair use. Please obtain copies from the publisher if appropriate.

1. Design criteria for engineering inorganic material-specific peptides.
   Langmuir 2005 21(15):6929-6933
   Peelle BR, Krauland EM, Wittrup KD, Belcher AM
   URL Image:Macintosh HD-Users-nkuldell-Desktop-BE109-BE109 lab manual S06-Mod4 (S06)-MetalBindingYeast Lang05.pdf Image:Macintosh HD-Users-nkuldell-Desktop-BE109-BE109 lab manual S06-Mod4 (S06)-MetalBindYeast Sup1 Lang05.pdf Image:Macintosh HD-Users-nkuldell-Desktop-BE109-BE109 lab manual S06-Mod4 (S06)-MetalBindYeast Sup2 Lang05.pdf