Difference between revisions of "Part:BBa I763019: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.
+
We cloned this 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 the same cloning in a low copy number plasmid (pSB4A3), thinking that it could better match with a small amount of endogenous cI repressor, but we obtained the same result.
  
As expected, we observed that bacteria transformed with BBa I763032 are no fluorescent in LB medium untill they reached the growth stationary phase. When all the glucose was depleted bacteria were fluorescent due to lack of the catabolite repression from cAMP- CAP complex.
+
As expected, we observed that bacteria transformed with BBa I763032 are not fluorescent in LB medium until they reached the growth stationary phase. When all the glucose was depleted bacteria were fluorescent due to lack of the catabolite repression from cAMP- CAP complex.
  
 
===Applications of BBa_I763019===
 
===Applications of BBa_I763019===

Revision as of 17:06, 26 October 2007


We cloned this 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 the same cloning in a low copy number plasmid (pSB4A3), thinking that it could better match with a small amount of endogenous cI repressor, but we obtained the same result.

As expected, we observed that bacteria transformed with BBa I763032 are not fluorescent in LB medium until they reached the growth stationary phase. When all the glucose was depleted bacteria were fluorescent due to lack of the catabolite repression from cAMP- CAP complex.

Applications of BBa_I763019

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