Difference between revisions of "Part:BBa K776030:Experience"
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<I>'''iGEM CINVESTAV_IPN_UNAM''' CHARACTERIZATION OF IGEM DISTRIBUTION BIOPARTS</I> | <I>'''iGEM CINVESTAV_IPN_UNAM''' CHARACTERIZATION OF IGEM DISTRIBUTION BIOPARTS</I> | ||
| − | For contribute to the parts registry our team decided to make the characterization of constitutive promoters belonging to the family isolated from a small combinatorial library (J23101 , J23102, J23104, J23107, J23108, J2311, and J23115) which were attached to GFP to determine promoter activity, using the equipment Victor X3 Multilabel Plate Reader. | + | For contribute to the parts registry our team decided to make the characterization of constitutive promoters, in ''E. coli'', belonging to the family isolated from a small combinatorial library (J23101 , J23102, J23104, J23107, J23108, J2311, and J23115) which were attached to GFP, in psB1C3, to determine promoter activity, using the equipment Victor X3 Multilabel Plate Reader. |
[[Image:Gfp1.jpg]] | [[Image:Gfp1.jpg]] | ||
Latest revision as of 18:53, 29 September 2012
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Applications of BBa_K776030
iGEM CINVESTAV_IPN_UNAM CHARACTERIZATION OF IGEM DISTRIBUTION BIOPARTS
For contribute to the parts registry our team decided to make the characterization of constitutive promoters, in E. coli, belonging to the family isolated from a small combinatorial library (J23101 , J23102, J23104, J23107, J23108, J2311, and J23115) which were attached to GFP, in psB1C3, to determine promoter activity, using the equipment Victor X3 Multilabel Plate Reader.
Fig. 1 Construction of the promoter J23107 expressing GFP.
Methods
With the selected colonies, an overnight culture was made in M9 media(minimal media supplemented with 0.2% CAA). After 12 hours the culture was transferred to a 96 well plate at a 1:10 dilution (20 μl of culture and 180 μL of fresh M9 medium). OD and fluorescence measurements of the selected colonies were performed at intervals of 30 minutes for 16 h. From the results the PopS were calculated (polymerases per second).
Modeling
The ecuations used for calulated de promoter activity were based on (R. K. Jason et. al 2009).
Results
In the following graphs there is shown the GFP expression in function of th time and the realtive promotor intensity.
With the previous results of the characterization of the promoters there is concluded that the promoter J23107, is the strongest because it produces more RPUs”
User Reviews
UNIQbee8ce866d0abb17-partinfo-00000000-QINU UNIQbee8ce866d0abb17-partinfo-00000001-QINU