Difference between revisions of "Part:BBa K2433004"

Revision as of 08:10, 1 November 2017

VirB1

The VirB1 promoter is an inducible promoter for virB1 that was taken from an Agrobacterium Tumefaciens plasmid. This promoter is induced by acetosyringone and VirB1 is involved in virulence in wild type A. Tumefaciens. Wounded plants release acetosyringone, causing A. Tumefaciens to up-regulate the expression of VirB1 when near a wound. In the context of synthetic biology, the VirB1 promoter is a useful tool to control gene expression. For example, in A. Tumefaciens the virB1 promoter could be used to keep a potentially toxic synthetic gene like CRISPR quiescent until the bacteria neared a plant wound. Functional virG and virA are also required for virB induction. VirG and virA are both weakly inducible by acetosyringone.

The VirB1 promoter was cloned into two backbones (K135010 and J04650) containing red fluorescent protein (RFP) and transformed into the Agrobacterium Tumefaciens strain GV3101 for further testing. This strain contains functional virG and virA

A growth assay was performed using cultured A. Tumefaciens containing the VirB1-RFP fusion in the J04650 backbone. Two overnight cultures were inoculated from the same colony and each culture was split into three tubes for further treatment. Two tubes from each culture were treated with 100uM acetosyringone while the third was used as an un-induced control. Microplate measurements were taken at 6.5 hours and 23.5 hours, with no significant difference observed between un-induced and induced samples. As such, this part is not experimentally verified as being induced by acetosyringone.

Future experiments are needed to validate whether this part is inducible by acetosyringone. The literature offers several explanations for the apparent lack of inducibility in the aforementioned assay. Notably, it has been found that

Sequence and Features