Device

Part:BBa_K1797002

Designed by: iGEM15_Tsinghua   Group: iGEM15_Tsinghua   (2015-09-17)

IPTG-inducible CcdB

This part includes lacI and lac operator which are intended for the control of the expression of CcdB. LacI is constitutively expressed and binds to lac operator to block the expression of CcdB. In the presence of IPTG, lacI is released from operator and CcdB can be expressed. CcdB is a poisonous protein for E.coli since it affects the activity of DNA gyrase and inhibits the segregation of chromosomal DNA. Thus, when IPTG is added,the bacteria containing this part will go through cell death.


Part Demonstration

To test this part, we added 0.2% IPTG of 0 microliter, 1 microliter, 2 microliter and so on until 26 microliter respectively to 3ml E.coli culture solution. After cultured at 37℃ for about 20 hours, we measured the OD600 value of each tube and made a figure. From the following figure we can find that with the increase of IPTG concentration, the OD600 value decreases correspondingly. This observation confirms the usefulness of our part.

Fig.1 OD600-IPTG concentration


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1208
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None