Difference between revisions of "Part:BBa K2114000"

Revision as of 16:03, 12 October 2016

PCotYZ-RBS

PCotYZ promoter from Bacillus subtilis with ribosome binding site.

Usage and Biology

The PCotYZ promoter from Bacillus subtilis plays an important role in the sporulation of B. subtilis. The Promoter is located in the cotVWXYZ gene cluster and drives the expression of the late-stage spore crust proteins CotY and CotZ[1]. We included the naturally occurring ribosome binding site for cotZ to the promoter while maintaining the BioBrick compatibility. The improved promoter can be used for the 3A assembly with a coding region and cloned into an integration vector. The resulting device enables the transformation of B. subtilis for integration and expression of desirable genes.

Cloning strategy of the promoter improvement. PCotYZ was amplified with primers containing extensions in order to introduce the ribosome binding site. The resulting fragment was digested with XbaI and PstI and ligated into the linearized pSB1C3 vector with the corresponding overhangs.

Characterization

Sequence and Features