Regulatory

Part:BBa_K1033220:Experience

Designed by: Stephanie Herman, Alona Nyberg   Group: iGEM13_Uppsala   (2013-08-23)


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_K1033220

Utah State 2015 iGEM Team Use

The Utah State 2015 iGEM Team used this, along with two other related promoters (CP8, CP11, CP44) in a Lactococcus lactis project. Due to time constraints and difficulties working with non-model organisms, the constructs were not inserted into L. lactis but were cloned into Escherichia coli and compared in strengths using the sfGFP(Bs) and mCherry(Lr) fluorescent proteins (see Figures 1 and 2, respectively). The findings were not consistent across different fluorescent proteins, which may be due to secondary structure differences in mRNA or in the protein sequences themselves. These data were very consistent for each of the six constructs over several biological and technical replicates.

Utah_State_2015_sfGFP%28Bs%29_Fluorescence_Chart_Version3.jpeg

Figure 1. Fluorescence levels from three constructs using synthetic promoters from Jensen and Hammer (1998) in E. coli excited at 485/20 with emissions read at 528/20. cp8 from part BBa_K1820014, cp11 from part BBa_K1820015, and cp44 from part BBa_K1820016.

Utah_State_2015_MCherry_Fluorescence_Chart_Version_2.jpg

Figure 2. Fluorescence levels from three constructs using synthetic promoters from Jensen and Hammer (1998) in E. coli excited at 530/25 with emissions read at 590/35. cp8 from part BBa_K1820017, cp11 from part BBa_K1820018, and cp44 from part BBa_K1820019

Referrence: Jensen, P. R., Hammer, K. (1998). The Sequence of Spacers between the Consensus Sequences Modulates the Strength of Prokaryotic Promoters. Appl Environ Microbiol. 1998 Jan; 64(1): 82–87. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC124675/

User Reviews

UNIQ67240265c5aaa3ba-partinfo-00000000-QINU

No review score entered. Username

This part was used in the composite part BBa_K1365555 for characterization by the Rijksuniversiteit Groningen.

Graphricksfgfpbs.png

The graph shown above helps us to know distribution of the fluorescence in the cells of E.coli with different CP promotor fused to superfoldedGFP-terminator-RBS collection (CP1, CP11, CP29, CP30, CP41, CP44) and J23101 as reference. We used this method to characterize the CP promoter collection that was BioBricked by the Uppsala 2013 team.

;

UNIQ67240265c5aaa3ba-partinfo-00000002-QINU