Difference between revisions of "Part:BBa K4307021"
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__NOTOC__ | __NOTOC__ | ||
| − | + | !--PnisRRH07 -- | |
| − | + | NOTOC__ | |
| + | partinfoBBa_K4307021 shortpartinfo | ||
| − | + | This part is derived from PnisA, which is originated from iLactococcus lactis subsp. Lactisi, by the improvement done by our own software tool to enhance the promoter. | |
| + | |||
| + | |||
| + | |||
| + | !-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | + | !-- -- | |
| − | + | span class='h3bb'Sequence and Featuresspan | |
| − | + | partinfoBBa_K4307021 SequenceAndFeaturespartinfo | |
| + | |||
| + | |||
| + | !-- Uncomment this to enable Functional Parameter display | ||
| + | ===Functional Parameters=== | ||
| + | partinfoBBa_K4307000 parameterspartinfo | ||
| + | !-- -- | ||
| + | |||
| + | |||
| + | |||
| + | |||
| + | h2Characterization h2 | ||
| + | |||
| + | pThe following figure demonstrates our successful construction.p | ||
| + | |||
| + | br | ||
| + | httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307021-gel.pngbr | ||
| + | bFigure 1 b bThe construction results of PnisRRH07.b | ||
| + | |||
| + | |||
| + | |||
| + | h3Fluorescence assay was done to characterize the bio brick. h3 | ||
| + | pWe introduced these 10 promoters into our system into MACH1-T1 strain to detect EGFP fluorescence signal with and without nisin induction by microplate reader. Results demonstrated that fluorescence signals were enhanced both with and without nisin induction except for PnisRRH07 and PnisRRH10(Figure 2). p | ||
| + | p | ||
| + | br | ||
| + | httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307021-1.pngbr | ||
| + | bFigure 2 b b Fluorescence spectrophotometry results of the 10 mutant promoters with the highest predictive strength.b | ||
| + | |||
| + | br | ||
| + | Furthermore, flow cytometry results confirmed that percentages of positive fluorescence signal bacteria also increased both with and without nisin induction(Figure 3), indicating that the improvement of promoter PnisA by our software tool was practical. | ||
| + | br | ||
| + | httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307015-2.pngbr | ||
| + | bFigure 3 b b Flow cytometry results of the 10 mutant promoters with the highest predictive strength. b | ||
| + | |||
| + | br | ||
| + | |||
| + | p | ||
| + | |||
| + | h2Conclusionh2 | ||
| + | pAccording to fluorescence assay result, it can be found that the improvement of promoter PnisA by our software tool is a great success, though the effectiveness of PnisRRH07 and PnisRRH10 is relatively low. This helps us a lot in providing an improve to the composite part J23100-nisK-nisR+PnisA-EGFP(BBa_K4307013) and create better nisin induced expression system in iE.colii.p | ||
| + | |||
Revision as of 16:03, 12 October 2022
!--PnisRRH07 --
NOTOC__ partinfoBBa_K4307021 shortpartinfo
This part is derived from PnisA, which is originated from iLactococcus lactis subsp. Lactisi, by the improvement done by our own software tool to enhance the promoter.
!-- Add more about the biology of this part here
Usage and Biology
!-- -- span class='h3bb'Sequence and Featuresspan partinfoBBa_K4307021 SequenceAndFeaturespartinfo
!-- Uncomment this to enable Functional Parameter display
Functional Parameters
partinfoBBa_K4307000 parameterspartinfo !-- --
h2Characterization h2
pThe following figure demonstrates our successful construction.p
br httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307021-gel.pngbr
bFigure 1 b bThe construction results of PnisRRH07.b
h3Fluorescence assay was done to characterize the bio brick. h3 pWe introduced these 10 promoters into our system into MACH1-T1 strain to detect EGFP fluorescence signal with and without nisin induction by microplate reader. Results demonstrated that fluorescence signals were enhanced both with and without nisin induction except for PnisRRH07 and PnisRRH10(Figure 2). p p
br
httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307021-1.pngbr
bFigure 2 b b Fluorescence spectrophotometry results of the 10 mutant promoters with the highest predictive strength.b
br Furthermore, flow cytometry results confirmed that percentages of positive fluorescence signal bacteria also increased both with and without nisin induction(Figure 3), indicating that the improvement of promoter PnisA by our software tool was practical. br httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307015-2.pngbr
bFigure 3 b b Flow cytometry results of the 10 mutant promoters with the highest predictive strength. b
br
p
h2Conclusionh2 pAccording to fluorescence assay result, it can be found that the improvement of promoter PnisA by our software tool is a great success, though the effectiveness of PnisRRH07 and PnisRRH10 is relatively low. This helps us a lot in providing an improve to the composite part J23100-nisK-nisR+PnisA-EGFP(BBa_K4307013) and create better nisin induced expression system in iE.colii.p