Part:BBa_K606046:Experience

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Re-cloning of KinA

We suceeded in recovering the KinA gene from the non biobricked plasmid synthetized de novo by the 2009 Newcastle team, and cloned it into a standard biobrick plasmid, pSB1C3. Then we cloned this gene in front of the pVeg-SpovG (K143051) promoter + RBS. These two constructs had been sended to the registry into pSB1C3.

Characterization of the sporulation induced by the expression of KinA

We used a strain holding a KinA gene under the control of a hyperspank promoter. Growing the cell into a synthetic minimal media for hours, we saw the cell starting sporulating under the microscope after 3h. Here are the images from the characterization.

Sporulation assay with KinA overexpression

Induction of sporulation after 3h on CDH Medium +4mM IPTG	Induction of sporulation after 3h on CDH Medium +4mM IPTG
Negative control of sporulation after 3h on CDH Medium +0mM IPTG

Ref: Single,chemically defined sporulation medium for bacillus subtilis: growth,sporulation,and extracellular protease production. James H.HAGEMAN et al

We suceeded in recovering the KinA gene from the non biobricked plasmid synthetized de novo by the 2009 Newcastle team, and cloned it into a standard biobrick plasmid, pSB1C3. Then we cloned this gene in front of the pVeg-SpovG (K143051) promoter + RBS. These two constructs had been sended to the registry into pSB1C3.

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