Difference between revisions of "Part:BBa R0011:Experience"
| Line 142: | Line 142: | ||
|} | |} | ||
| + | |||
| + | |||
| + | {|width='80%' style='border:1px solid gray' | ||
| + | |- | ||
| + | |width='10%'| | ||
| + | <partinfo>BBa_K592008 AddReview 5</partinfo> | ||
| + | <I>[http://2013.igem.org/Team:UCSF UCSF 2013]</I> | ||
| + | |width='60%' valign='top'| | ||
| + | |||
| + | <html> | ||
| + | <regulartext1> The UCSF iGEM team tested the plAC promoter (part BBa_R0011) by placing it in front of GFP and inducing with various concentrations of IPTG. We used this part in the design of our CRISPRi circuit and improved upon it through the following experiments. <br> | ||
| + | |||
| + | <img align="center" style="margin-bottom:0px; width: 500px; margin-top:20px; padding:2; margin-left:55px;" src="https://static.igem.org/mediawiki/2013/9/9a/PLAC_dose_response.png"> <br> | ||
| + | |||
<!-- DON'T DELETE --><partinfo>BBa_R0011 EndReviews</partinfo> | <!-- DON'T DELETE --><partinfo>BBa_R0011 EndReviews</partinfo> | ||
Revision as of 17:35, 27 September 2013
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_R0011
User Reviews
UNIQ1a4f7b3968e51802-partinfo-00000000-QINU
|
•••••
Antiquity |
This review comes from the old result system and indicates that this part worked in some test. |
|
UNIPV-Pavia iGEM 2009 |
UNIPV-Pavia iGEM 2009's Experience: BBa_R0011 - Plac hybrid promoter |
|
•••
UNIPV-Pavia iGEM 2010 |
|
Washington 2010 |
Washington 2010's Experience: R0011 in different plasmids |
|
••••
iGEM Kyoto 2010 |
We measured RPUs of R0011 in low number copy plasmid with various concentrations of IPTG |
|
••••
iGEM11_Uppsala-Sweden |
We measured RPUs of different promoters in low copy number plasmids. |
|
•••••
UT-Tokyo 2011 |
UT-Tokyo 2011 Team has characterized this part through the test of our Firefly-Renilla Dual Luciferase Assay Kit. We succesfully evaluated the relative expression levels of BBa_R0011 with various IPTG concentration, compared to that of BBa_J23119 as a control. For experimental details, see [http://2011.igem.org/Team:UT-Tokyo our page]. |
|
•••••
[http://2012.igem.org/Team:Uppsala_University Uppsala University 2012] |
iGEM Team Uppsala University 2012 Promoter strength A promoter test was carried out to put synthetic and natural promoters on the same scale. Every promoter was assembled before B0032-SYFP2 (BBa_K864101) in the backbone BBa_K592200 (very similar to the pSB3x5 backbones). The test was performed in E coli expression strain MG1655 and cloning strain DH5alpha, by flow cytometry fluorescence measurements of single cells. Triplicates of each strain and promoter were inoculated in 2 mL LB media with spectinomycin (50 µg/mL) and grown overnight shaking at 37° C. Samples were equilibrated in PBS solution at 1:160 dilution for one hour, and then measured by a BD Biosciences FACSaria III. 10^5 cells of each sample were individually measured and averaged, with dead and other non-flourescent cells excluded. Promoter strength is noted as fractions of the reference promoter's, J23101, strength in corresponding strain.
The variance in expression between MG1655 and DH5α may depend on the reference J23101. The maximum protein expression may be lower in DH5α, due to its lower fitness resulting in lower expression of SYFP2 in the J23101 construct. Alternatively, the clone with J23101 in DH5α may have been weaker than average, resulting in higher RPU values compared to other DH5α. LacI repression The possibility of repression by lacI and induction by IPTG was evaluated in the pSB4C15Iq backbone. Cells grown with IPTG (0.5 mM) had a 100-fold increase of RFP expression, when compared to those grown without. Read about pSB4C15Iq for details. |
|
•••••
[http://2013.igem.org/Team:UCSF UCSF 2013] |
|
