Yale 2010

1. Yale2010 Toward Bacterially Deposited Circuits 7-1-2010

2. Recent Wetlab Work • TSI agar assay of pSB74 containing transformants • Cloning work – PCR amplification of phsABC and insertion vector containing iGEM terminator phsABC B0015 phsABC B0015 phsABC B0015

3. Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu H2S H2S H2S H2S Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Extracellular Bound Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu H2S H2S H2S Cu H2S Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Cu Electric current CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS CuS

4. Extracellular Secretion Cu Cu Cu Cu Cu Cu CuS Cu H2S H2S CuS Cu Cu CuS H2S H2S Cu Cu Cu Cu CuS Cu Cu H2S Cu Cu Cu Cu Cu Cu Cu CuS CuS CuS CuS CuS CuS CuS CuS Secretion Signal on N-terminus and Avadin tag (or Strep tag) on C-terminus so the protein will bind to the biotin chip Biotin chip

5. Avidin or Strep-tag biotin

7. Pseudoazurin Thioredoxin J Biochem. 1994 Aug;116(2):327-34. Extracellular transport of pseudoazurin of Alcaligenes faecalis in Escherichia coli using the COOH-terminal domain of Serratia marcescens serine protease. J Biol Chem. 1996 Mar 1;271(9):5059-65. Histidine patch thioredoxins. Mutant forms of thioredoxin with metal chelating affinity that provide for convenient purifications of thioredoxin fusion proteins.

8. Examples of the secretion of recombinant proteins expressed as fusions to the HlyA signal sequence *Endochitinase (Trichoderma harzianum), green fluorescent protein (Aequorea victoria), human erythropoetin, and trout growth hormone. http://wolfson.huji.ac.il/expression/local/bacteria/Recombinant%20protein%20secretion%20in%20Escherichia%20coli-2005.pdfz