Types | DnaRegion
|
Roles | Composite
engineered_region
|
Sequences | BBa_K777104_sequence (Version 1)
|
Description
[[Image:Table_K777101-K777108_new.jpg|thumb|240px|right|Fig. 1: Overview of BioBricks containing the flhDC operon downstream of Anderson promoters. (Activity according to Berkeley 2006)]]
The flhDC operon is the master regulator of motility and chemotaxis in E. coli. FlhDC forms heterotetramers and activates class II operons in concert with sigma factor 70. Among the gene products of class II operons are several components of the flagellum and the alternative sigma factor FliA which is essential for the transcription of class III genes.
Here we used a set of 8 constitutive Anderson-promoters from the 2006 Berkeley group to test how different levels of constitutive flhDC expression affect the motility of E. coli.
Notes
- The PstI restriction site in the flhDC sequence was mutated to CTGCGG using the reverse primer
- flhDC was amplified from genomic DNA of E. coli DH10B via PCR using the following primers:
- Fwd: 5'-actgaattcgcggccgcttctagatgcatacctccgagttgctg-3'
- Rev: 5'-tcctgcagcggccgctactagttactgcccgggatggcggttgacataagcCgcaggcaaagctgccaacag-3'
(the capitalized "C" induces the mutation for removal of the PstI site)
Source
- The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B, complete genome (CP000948.1).
- J23106 information was taken from the partsregistry and physical DNA from the 2012 distribution kit.