Difference between revisions of "Part:BBa I712004"

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[[Image:CMV_standard2.png|center|400px|thumb|'''Figure 1: Strength of the CMV promoter in different cell lines'''. [http://2009.igem.org/Team:Heidelberg/Project_Measurement Relative Expression Units (REU)] was determined by Flow cytometry; in HeLa and MCF-7 cells, this measurement was confirmed by an independent technique (Image analysis). 3 experiments on different days, three replicates for each measurement.]]
 
[[Image:CMV_standard2.png|center|400px|thumb|'''Figure 1: Strength of the CMV promoter in different cell lines'''. [http://2009.igem.org/Team:Heidelberg/Project_Measurement Relative Expression Units (REU)] was determined by Flow cytometry; in HeLa and MCF-7 cells, this measurement was confirmed by an independent technique (Image analysis). 3 experiments on different days, three replicates for each measurement.]]
 
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[[Image:HD09_FACS_cmv_medium_with_time.PNG|center|400px|thumb| '''Figure 2: CMV strength changes depending on conditions''' As [http://2009.igem.org/Team:Heidelberg/Project_Measurement#There_are_no_truly_constitutive_promoters_in_mammalian_cells there are no truly constitutive promoters in mammalian cells], both the reference promoter ([[Part:BBa_K203112]]) and CMV fluctuate dependent on condition (Erolimus induces extreme starvation). Measured by Flow cytometry 20 ours after transfection (unless specified otherwise).]]
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[[Image:HD09_FACS_cmv_medium_with_time.PNG|center|400px|thumb| '''Figure 2: CMV strength changes depending on conditions''' As [http://2009.igem.org/Team:Heidelberg/Project_Measurement#There_are_no_truly_constitutive_promoters_in_mammalian_cells there are no truly constitutive promoters in mammalian cells], both the reference promoter ([[Part:BBa_K203112]]) and CMV fluctuate dependent on condition (Erolimus induces extreme starvation). Measured by Flow cytometry 20 hours after transfection (unless specified otherwise).]]
 
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Revision as of 15:56, 16 October 2009

CMV promoter

a constitutive expression promoter for use in mammalian cells. Ribosome binding site is included.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Functional Parameters

negative_regulators-NA-
positive_regulators-NA-

[http://2009.igem.org/Team:Heidelberg Heidelberg 2009 iGEM team] characterized this promoter in Part:BBa_K203100 as a first application for its newly developed units of promoter strength in mammalian cells, [http://2009.igem.org/Team:Heidelberg/Project_Measurement Relative Expression Units (REU)] and [http://2009.igem.org/Team:Heidelberg/Project_Measurement Relative Mammalian Promoter Units (RMPU)], where RMPU is directly proportional to PoPs and measured on a RNA level, whereas REU is measured on the protein level. We found CMV to have a strength of 5,52 REU (Standard Error of the Mean = 0,60) in [http://2009.igem.org/Team:Heidelberg/Eucaryopedia#HeLa HeLa cells], 6,57 (SEM = 0,91) in [http://2009.igem.org/Team:Heidelberg/Eucaryopedia#MCF-7 MCF-7 cells] and 9,96 (SEM = 1,52) in [http://2009.igem.org/Team:Heidelberg/Eucaryopedia#U2-OS U2-OS cells] cells (Fig 1). We furthermore found that CMV is not a truly constitutive promoter (but we [http://2009.igem.org/Team:Heidelberg/Project_Measurement#There_are_no_truly_constitutive_promoters_in_mammalian_cells argue] that no such thing exists. In RMPU, CMV has an activity of approx. 2.89 RPMU in HeLa cells (Figures to be added).

Figure 1: Strength of the CMV promoter in different cell lines. [http://2009.igem.org/Team:Heidelberg/Project_Measurement Relative Expression Units (REU)] was determined by Flow cytometry; in HeLa and MCF-7 cells, this measurement was confirmed by an independent technique (Image analysis). 3 experiments on different days, three replicates for each measurement.
Figure 2: CMV strength changes depending on conditions As [http://2009.igem.org/Team:Heidelberg/Project_Measurement#There_are_no_truly_constitutive_promoters_in_mammalian_cells there are no truly constitutive promoters in mammalian cells], both the reference promoter (Part:BBa_K203112) and CMV fluctuate dependent on condition (Erolimus induces extreme starvation). Measured by Flow cytometry 20 hours after transfection (unless specified otherwise).