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.

S. cerevisiae shown budding (top) and bound to CdS (bottom)

Lab handouts

Day 1: Getting to know yeast

Day 2: Panning for gold binders

Day 3: Screening library and optimizing panning

Day 4: Checking gold binders

Day 5: PCR of gold binding candidates

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 PDF reprint first supplement to pdf second supplement to pdf