Reporter

Part:BBa_K1223007:Design

Designed by: Orr Schlesinger   Group: iGEM13_BGU_Israel   (2013-09-07)
Revision as of 13:12, 12 October 2013 by Alex m (Talk | contribs)

cI translational unit with His-tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is an improvement of the existing BBa_K327018 LVA tagged cI repressor protein. We added a his tag instead of the LVA tag in order to offer a convenient way to study the protein without damaging it. The his tag is located at its C-terminal, therefore having no effect on its function.

we used this part in order to identify our protein through western blot technique, using anti His-Tag antibodies. in the western it is apparent that without induction there is no leakage of the cI protein but upon induction we get a nice band indicating that the cI is expressed. The positive control we used was a model protein containing a His tag.

CIwestern.jpg


Source

part is synthetic. the cI CDS (Originated from the lambda bacteriophage) and the promoter were taken from the iGEM registry.

References

M. Pedersen, M. Ligowska, K. Hammer, Characterization of the CI repressor protein encoded by the temperate lactococcal phage, Journal of Bacteriology 29, [2010]