Our team has contributed to the iGEM community through the development of several parts collections and the characterization and improvement of existing parts.
The first collection contains parts for use in the creation of an acid-sensing, base-producing genetic circuit, and a base-sensing, acid-producing circuit. These parts were created to test the function of the pH genetic elements, and to create a device that can sense the pH of wastewater from manufacturing and respond by producing acid or base to neutralize the waste. Information about these parts can be found in our Engineering Success page.
The second collection was created to test what effect pH might have on the excitation and emissions spectra of fluorescent proteins. As part of our project, we utilized fluorescent proteins in reporter constructs to test pH responsive change in gene expression. We realized we had no information on what effect pH might have on the fluorescent proteins used as reporters. We therefore created genetic constructs that we could use to purify BFP and RFP, so we could investigate this. More information about this work can be found on our Measurement page.
| Part number | Part name | Function | Description |
|---|---|---|---|
| BBa_K5097000 | PRE | Regulatory | pH Response Element riboswitch |
| BBa_K5097001 | PREmR34 | Reulatory | pH Response Element riboswitch mR34 |
| BBa_K5097002 | PREm13 | Regulatory | pH Response Element riboswitch m13 |
| BBa_K5097003 | PRE-BFP | Composite | PRE-BFP reporter |
| BBa_K5097004 | PREmR34-BFP | Composite | PREmR34-BFP reporter |
| BBa_K5097005 | PREm13-BFP | Composite | PREm13-BFP reporter |
| BBa_K5097006 | PREm13-LDHA | Composite | Base-sensor, acid-producer PREm13-LHDA |
| BBa_K5097008 | pASR-RFP | Composite | pASR-RFP reporter |
| BBa_K5097009 | SpeA | Coding | SpeA (arginine decarboxylase) |
| BBa_K5097010 | SpeB | Coding | SpeB (Agmatinase) |
| BBa_K5097011 | pASR-PUT1 | Composite | Acid-sensor, Base-producer pASR-PUT1 |
| BBa_K5097012 | RBS with spacer | Regulatory | RBS BBa_B0034 with spacer nucleotides |
| BBa_K5097013 | 6X His tag with spacer | Tag | 6X His tag with AGVG linker |
| BBa_K5097014 | sBFP without a stop codon | Coding | sBFP without a stop codon |
| BBa_K5097015 | RFP without a stop codon | Coding | RFP without a stop codon |
| BBa_K5097016 | sBFP with 6X His | Composite | sBFP with 6X His tag |
| BBa_K5097017 | RFP with 6X His | Composite | RFP with 6X His tag |
Table 1: Summary table describing the pHish and CHIPS parts collections
Our second contribution came about through our development of a measurement study to characterize the pH-dependent signals of commonly used fluorescent reporters from the iGEM Parts Repository. This latter work is described on our Measurement page.