Question

If you were given access to the Biobrick collection of biological parts and asked to construct an artificial circuit capable of producing two stable states of gene expression, describe

1)    the types of parts you would choose and how you would arrange them [6 min]

2)    how you would demonstrate experimentally that your circuit behaved as planned. [4 min]

Answer 1

1) To construct an artificial circuit, I would use 9 main parts which represents a general circuit for the production of two stable states of gene expression:

1)Cloning site prefix and suffix

2)Positive selection marker

3)Replication origin

4)Translational stop sequence

5)Forward transcriptional terminator

6)Reverse transcriptional terminator

7)Resistance marker

8)Forward sequencing primer annealing

9)Reverse sequencing primer annealing site

10)Restriction site

Replication origin

2) To demonstrate experimentally that my circuit behaved as planned, I would use an Elisa Assay to detect levels of the proteins that I'm trying to replicate using the circuit. This will be done using specific antibodies against the target proteins.

Positive Cloning site Cloning Replicatiorn origin selection marker suffix site prefix Translational Translational stop sequence stop sequence Reverse Forward transcriptional transcriptional terminator terminator Forward Reverse transcriptional transcriptional terminator terminator Resistance Replication origin marker