Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: The Newcastle 2009 iGEM team
Date created: 2009-10-12 11:00:00
Date modified: 2015-05-08 01:10:58
IPTG inducable kinA Bacillus sporulation trigger
| Types | DnaRegion |
| Roles | engineered_region Device |
| Sequences | BBa_K174011_sequence (Version 1) |
Description
Spo0A is the master regulator of sporulation in B. subtilis. When it is phosphorylated, it activates the expression of more than 100 genes(1). It has been shown that, activation of spo0A is sufficient to trigger sporulation.By artifically synthesising KinA during growth, spoulation can be triggered in a multicomponent phosphorelay in which KinA phosphorylates Spo0F, Spo0F~P in turn phosphorylates Spo0B, and finally Spo0B~P phosphorylates Spo0A. (2)
Hence by adding pspac promoter and an RBS to the front of kinA coding sequence, we made a device that can express KinA when induced with IPTG. Hence we can trigger sporulation by adding IPTG to the medium.
Spo0A~ P initiates sporulation first by activating low-threshold genes and then by gradually increasing and activating high-threshold genes. Hence the multicomponent phosphorelay???s elements, KinA, Spo0F and spo0B, gradually increases spo0A and its activity, and initiates the sporulation(2).
Notes
In B. subtilis, out of five kinases only KinA, KinB and KinC kinases are effective to trigger the sporulation. KinD and KinE do not phosphorylates Spo0F, hence do not effect the sporulation. We chose KinA for our device.Source
Synthesised.| Sequence Annotation | Location | Component / Role(s) |
| AccIII Cut Site BBa_K174004 pspac promoter BalI Cut Site RBS BBa_K174010 KinA | 1,6 7,112 113,118 119,135 136,1953 | sequence_feature feature/dna feature/promoter promoter feature/dna sequence_feature ribosome_entry_site feature/rbs feature/BioBrick engineered_region |