Difference between revisions of "Part:BBa I712004"
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− | [[Image:QPCR_result_matlab.png|thumb|center|350px|<div style="text-align:justify;">'''Fig. | + | [[Image:QPCR_result_matlab.png|thumb|center|350px|<div style="text-align:justify;">'''Fig. 3: Real-time RT-PCR data of CMV and JeT promoters.'''One group of HeLa cells were transfected with plasmid containing CMV promoter coupled to GFP. Another group with JeT promoter coupled to GFP was used as reference. RNA was extracted after 20 h and 50 h, followed by real-time RT-PCR. The Ct values were collected with a threshold of 0.05. The CMV activity compared to JeT at the same time point was calculated in MatLab as "arbitrary units" which correspond to amount of mRNA.</div> ]] |
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− | [[Image:qpcr_result.png|thumb|center|300px|<div style="text-align:justify;">''Fig. | + | [[Image:qpcr_result.png|thumb|center|300px|<div style="text-align:justify;">'''Fig. 4: Real-time RT-PCR data of CMV promoter''' Arbitrary units of CMV divided by that of JeT is the [http://2009.igem.org/Team:Heidelberg/Project_Measurement RMPU].</div>]] |
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Revision as of 20:38, 21 October 2009
CMV promoter
a constitutive expression promoter for use in mammalian cells. Ribosome binding site is included.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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 promoter exists). In [http://2009.igem.org/Team:Heidelberg/Project_Measurement RMPU] (a mRNA-based unit directly proportional to PoPs), CMV has an activity of approx. 2.89 RPMU in HeLa cells,as measured by qRT-PCR (Fig. 3)