Difference between revisions of "Part:BBa K606046:Experience"
 
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<h2>Re-cloning of KinA</h2>
 
<h2>Re-cloning of KinA</h2>
 
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.
 
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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<h2>Characterization of the sporulation induced by the expression of KinA</h2>
 
<h2>Characterization of the sporulation induced by the expression of KinA</h2>
  
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<p>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 soprulating under the microscope after 1h30. Here are the images from the characterization.</p>
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<p>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.</p>
  
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{| border="1" class="wikitable" style="text-align: center;"
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|+Sporulation assay with KinA overexpression
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|[[Image:spor1.jpg|450px|thumb|center|Induction of sporulation after 3h on CDH Medium +4mM IPTG]]
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|[[Image:spor2.jpg|450px|thumb|center|Induction of sporulation after 3h on CDH Medium +4mM IPTG]]
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|[[Image:control spor.jpg|450px|thumb|center|Negative control of sporulation after 3h on CDH Medium +0mM IPTG]]
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<img src="https://static.igem.org/mediawiki/parts/6/68/Spor1.jpg">
 
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<img src="https://static.igem.org/mediawiki/parts/5/51/Spor2.jpg">
 
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<img src="https://static.igem.org/mediawiki/parts/7/7a/Control_spor.jpg">
 
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Latest revision as of 04:50, 22 September 2011

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Applications of BBa_K606046

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.

===User Reviews=== BBa_K606046 StartReviews BBa_K606046 EndReviews