Difference between revisions of "Part:BBa K1088026"

Revision as of 18:38, 19 September 2013

B. subtilis dxs-GFP protein fusion (lac promoter with lac inhibitor: IPTG inducible)

This part consist of the dxs gene derived from B. subtilis fused to GFP at the translational level with a 10 AA linker between the proteins. The reporter fusion is under the control of the lac promoter and has a strong RBS.

To repress expression from the lac promoter, the lacI gene under a constitutive promoter, with a strong RBS and a efficient terminator is placed counter-clockwise to the reporter fusion. The repression can then be relieved with addition of IPTG, which binds and inhibits the function of LacI.

The levels of expression was meassured in E. coli K-12 MG1655 using fluorescence activated cell sorting (FACS), and proved that their was no expression when grown without IPTG and that there was expression after addition of IPTG. See experience for more details.

This Brick was build to test induction time and IPTG concentration for induction of a similar device (BBa_K1088027) which lacks the linker and GFP.

Sequence and Features