Difference between revisions of "Part:BBa I763032:Experience"

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We cloned the part inside a high copy number plasmid (pSB1AK3. When we checked part lenght with a gel electrophoresis that was ok. Anyway, when we checked for fluorescence we observed that just some bacteria were fluorescent. We grew bacteria in M9 + lactose + IPTG (2 mM),but always we saw that not all of them were fluorescent.
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As expected, we observe that bacteria transformed with BBa I763032 are no fluorescent in LB medium untill they reach the growth stationary phase. When all the glucose is depleted bacteria are fluorescent due to lack of the catabolite repression from cAMP- CAP complex.
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We first cloned the part inside a high copy number plasmid (pSB1AK3). When we checked part lenght with a gel electrophoresis that was ok. Anyway, when we checked for fluorescence we observed that just some bacteria were fluorescent. We grew bacteria in M9 + lactose + IPTG (2 mM),but always we saw that not all of them were fluorescent.
  
 
So, we tried tried the same cloning in a low copy number plasmid (pSB4A3), thinking that it could better with a lower amount of cI repressor, but we obtained the same result.
 
So, we tried tried the same cloning in a low copy number plasmid (pSB4A3), thinking that it could better with a lower amount of cI repressor, but we obtained the same result.

Revision as of 13:28, 26 October 2007


As expected, we observe that bacteria transformed with BBa I763032 are no fluorescent in LB medium untill they reach the growth stationary phase. When all the glucose is depleted bacteria are fluorescent due to lack of the catabolite repression from cAMP- CAP complex.

We first cloned the part inside a high copy number plasmid (pSB1AK3). When we checked part lenght with a gel electrophoresis that was ok. Anyway, when we checked for fluorescence we observed that just some bacteria were fluorescent. We grew bacteria in M9 + lactose + IPTG (2 mM),but always we saw that not all of them were fluorescent.

So, we tried tried the same cloning in a low copy number plasmid (pSB4A3), thinking that it could better with a lower amount of cI repressor, but we obtained the same result.


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