"

From 2011.igem.org

Contents 1 Verification of ΔcheZ strain (protocol) 2 Verification of the function of the constructed riboswitch (protocol) 3 Verification of Toggle-switch (protocol) 4 Modulate Toggle-switches to produce more balanced ratio (protocol) 5 Test of Incorporated Riboswitch into one side of Toggle-switch 6 Verification of the constructed system’s function

Verification of ΔcheZ strain (protocol)

The size of colonies of RP1616 was much smaller than that of RP437 under same circumstances and after same period of incubation time. In colony PCR following, RP437 absolutely has much more outcome than RP1616 and we concluded that RP1616 is a ΔcheZ strain.

Verification of the function of the constructed riboswitch (protocol)

1. 0.25% Semisolid medium
   
   
2. 0.4% Semisolid medium
   
   
3. 0.3% Semisolid medium
   
   

    From the results above we concluded that the constructed riboswitch functions well, especially on 0.3% Semisolid medium.

Verification of Toggle-switch (protocol)

                        

Numbers of colonies with RFP: Numbers of colonies with GFP ≈ 8:25

Modulate Toggle-switches to produce more balanced ratio (protocol)

                             

Numbers of colonies with RFP: Numbers of colonies with GFP ≈ 6:1

Test of Incorporated Riboswitch into one side of Toggle-switch

                       

Verification of the constructed system’s function

• 6 tubes of constructed monoclonal RP1616

• Results of the sixth tube