Part:BBa_K1731000:Experience

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Applications of BBa_K1731000

As a part of a TCS (Two Component System), this chimera can be used to design and test a circuit involving a great variety of signals and the expression of many genes. It could also be used in order to test positive and negative feedback pathways.

As a proof of concept, this part could be useful to design other types of possible functional chimeras.

User Reviews

UNIQcdd79e7b1e8b3b60-partinfo-00000000-QINU UNIQcdd79e7b1e8b3b60-partinfo-00000001-QINU

Two component systems are the major signal transduction mechanisms in bacteria, several receptors can sense some compounds in the media to respond with chemotaxis and metabolic changes. There are several periplasmic binding proteins that are the first sensors to a broad range of molecules, one of our interest is Glucose-Galactose Binding Protein (GGBP)(Dwyer and Hellinga 2004). Glucose is sensed by bacteria through the GGBP which has been proposed as a glucose sensor (Hsieh et al. 2004). Trg-EnvZ was initially created in 1994, in order to measure the carbohydrates concentration and using the EnvZ intracellular with a reporter, Trg chemoreceptor triggers the chemotaxis of the bacteria while the EnvZ is an osmoregulator which activates the transcription factor OmpR. This chimera allows an easy way to measure glucose concentration using a reporter for EnvZ(Tolosa and Rao 2006).

This part is aimed to work in order to achieve a bacterial glucose sensor coupled with insulin production as a response, the first biobrick (TrgZ) is designed to work as a glucose sensor which uses the intercellular domain of Trg that binds GGBP bound to glucose and activates the intracellular domain of EnvZ, this allow us to have a fine response to the glucose stimulus. The second part (Constitutive insulin lispro) is aimed to produce insulin in two modified E. coli strains (Rossettagami and SHuffle).

We have achieved to produce insulin in these two strains; we look forward to integrate the insulin production under the EnvZ stimulus and We will look forward to modulate the GGBP's glucose affinity by means of punctual mutations in order to adjust it to detect concentrations near human's physiological levels.