Results

Parts created

We created two new parts, [http://partsregistry.org/Part:BBa_K736000 BBa_K736000] (which regulated the transcription of downstream genes) and [http://partsregistry.org/Part:BBa_K736001 BBa_K736001] (which was a TAT signal sequence fused to a red fluorescent protein, E1010). We also assembled two parts, [http://partsregistry.org/wiki/index.php?title=Part:BBa_K736002 BBa_K736002] (which tests the ability of glucose to regulate transcription and uses enhanced RFP as the reporter) and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K736003 BBa_K736003] (which tests for the ability of the TAT sequence to cause secretion of protein outside of the cell and also uses RFP as a reporter). We also created one part that coded for human insulin chain A ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K736004 BBa_K736004]) and one part that coded for human insulin chain B ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K736005 BBa_K736005]). We are also submitting the DNA for [http://partsregistry.org/Part:BBa_K331009 K331009] which was in its planning stage in the registry.

Name	Type	Description	Length
[http://partsregistry.org/Part:BBa_K736000 BBa_K736000]	Regulatory	Constitutive Promoter (J23100) with MLC (glucose-regulated) binding site	36 bp
[http://partsregistry.org/Part:BBa_K736001 BBa_K736001]	Reporter	TAT signal sequence fused to red fluorescent protein (E1010)	720 bp
[http://partsregistry.org/Part:BBa_K736002 BBa_K736002]	Composite	Constitutive Promoter with MLC-binding-site-regulated enhanced RFP	913 bp
[http://partsregistry.org/Part:BBa_K736003 BBa_K736003]	Composite	pLAC-regulated TAT-RFP construct	948 bp
[http://partsregistry.org/Part:BBa_K736004 BBa_K736004]	Coding	Coding sequence for insulin chain A	72 bp
[http://partsregistry.org/Part:BBa_K736005 BBa_K736005]	Coding	Coding sequence for insulin chain B	99 bp

Proof of Principle

TAT signal sequence

RFP with Cells Media at Different Induction Levels

Cultures were grown containing pLacI promoter attached to a Twin Arginine tag signal sequence and fused to red fluorescent protein. Samples were taken at various time points and examined for fluorescents when excited at 586nm with an expected emission at 607nm. We used a fluorescents spectrophotometer to measure the red fluorescent protein present in the supernatant after spinning the cells to a pellet.

Glucose Variable Concentrations