Team:Evry/Chelator

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Iron coli project

Iron Chelator

First strategy for enterobactin biosynthesis

Here we present Fig 1 and 2 our constructions which contain each three Lac I regulated enterobactin synthesis genes. Escherichia coli naturally have those genes into a single operon but due to their important lenghts, we decided to divide them into two indivudual constructions in order to make the cloning easier.

Fig 1 First construction containing the Lac I regulated enterobactin synthesis genes Ent A, Ent D and Ent F. Genes fusions were made with flanking restriction sites that are compatible with Biobrick-based cloning.


Fig 2 Second construction containing the Lac I regulated enterobactin synthesis genes Ent B, Ent C and Ent E. Genes fusions were made with flanking restriction sites that are compatible with Biobrick-based cloning.

NAME FIGURE Description

Lac promoter

Lac Promoter

RBS + EntA

First gene required for enterobactin sythesis

RBS + EntB

Second gene required for enterobactin sythesis

RBS + EntC

Third gene required for enterobactin sythesis

RBS + EntD

Fourth gene required for enterobactin sythesis

RBS + EntE

Fifth gene required for enterobactin sythesis

RBS + EntF

Sixth gene required for enterobactin synthesis

Terminator

Transcription Stop signal

Plasmid

Backbone with ampicillin resistance

Table 1. Genetic elements used to produce the enterobactin siderophore.

Second strategy for enterobactin biosynthesis

HCversion
Figure 1: Légende ici.

Future caracterisation of the construction

Even though we tried to simplify the cloning, we did not succeed to obtain our two constructions. Thus, we have conceived new cloning approaches.