Logic gate overview
A functioning set of Locks and Keys and efficient cellular conjugation could lead to construction of basic [logic gates] in cells, which may be strung together to perform complex computation. We are desiging AND gates, and NAND gates. For a typical explanation of these gates, read up some wikipedia: AND gate and [NAND gate].
Design Phase 1: basic parts without conjugation or locks and keys(J23039, J23040, T9002, I13601).
Phase 1 was meant to use parts that "already work" to build a few gates and compare experiments and simulation.
An AND gate would be built with parts J23039 and T9002 <picture, and gate parts> J23039 encodes LuxI, which in turn creates the small molecule Homo-Serine Lactone. T9002 is GFP under Lux promoter, with LuxR present. LuxR inhibits the Lux promoter, but LuxI preferentially binds to Lux promoter. When LuxI is present, the Lux promoter is an effective promoter. So, T9002 recieves signal from J23039.
A NAND gate would be built with parts J23039, J23040, and I13601. <picture, parts> J23039 still encodes LuxI. J23040 is a LuxR/Lux promoter system (like T9002) that codes for LacI. LacI is an inhibitor. I13601 uses a Lac promoter, and codes for CFP. When LacI is present in the I13601, expression is off. When LacI is not present, expression is on. So, When J23039 and J23040 are present, I13601 signals "off," and NAND gate activity is created.
The plan for both of these parts. First, the parts would be built. Then, since the regulation we are using is a small molecule (homo-serine lactone made by LuxI) the parts would be tested without implementing conjugation. Finally, all of the parts would be put on plasmids with OriT, so that they could be conjugated into cells.
Design Phase 2: parts transferred via conjugation (when succesful, the first instance of actual gate activity)
Design Phase 3: parts under lock and key (when succesful, the first chance of stringing together gates)