Difference between revisions of "Part:BBa K1804001"
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<partinfo>BBa_K1804001 short</partinfo> | <partinfo>BBa_K1804001 short</partinfo> | ||
− | Green fluorescence protein (GFP) from ''Aequeora victoria'' was placed under the control of the constitutive promoter lacI ( | + | Green fluorescence protein (GFP) from ''Aequeora victoria'' was placed under the control of the constitutive promoter ''lacI'' (p''lac''). |
− | '''p''lac'' promoter is constitutively active in E. coli BL21''' | + | '''p''lac'' promoter is constitutively active in ''E. coli'' BL21''' |
− | The p''lac'' promoter is constitutively active in our strain of E. coli BL21. After construction of the pSB1C3-p''lac''-''gfp'' vector, the activity of the p''lac'' promoter was measured by quantification of fluorescence intensity. BL21 was used as the negative control and BL21 transformed with pAC-p''lac''-''gfp'' [[https://parts.igem.org/Part:BBa_K1804001:Design]] as the positive control. As seen in Figures 1 and 2, GFP is expressed in both strains of BL21 containing the p''lac''-''gfp'' vectors but not in the negative control. When fluorescence data was quantified in arbitrary units (A.U.), the intensity of p''lac''-''gfp'' in pSB1C3 was lower relative to that of p''lac''-''gfp'' in the pAC vector (Figure 2). Nonetheless, our results indicated that p''lac'' is indeed able to drive gene expression under normal growth conditions. Supplementary Table 1 shows the descriptive statistics of the fluorescence intensities determined. | + | The p''lac'' promoter is constitutively active in our strain of ''E. coli'' BL21. After construction of the pSB1C3-p''lac''-''gfp'' vector, the activity of the p''lac'' promoter was measured by quantification of fluorescence intensity. BL21 was used as the negative control and BL21 transformed with pAC-p''lac''-''gfp'' [[https://parts.igem.org/Part:BBa_K1804001:Design]] as the positive control. As seen in Figures 1 and 2, GFP is expressed in both strains of BL21 containing the p''lac''-''gfp'' vectors but not in the negative control. When fluorescence data was quantified in arbitrary units (A.U.), the intensity of p''lac''-''gfp'' in pSB1C3 was lower relative to that of p''lac''-''gfp'' in the pAC vector (Figure 2). Nonetheless, our results indicated that p''lac'' is indeed able to drive gene expression under normal growth conditions. Supplementary Table 1 shows the descriptive statistics of the fluorescence intensities determined. |
[[File:BBa_K1804001_Fluorescence.png|600px|thumb|center|'''Figure 1:''' Representative images of bacterial cultures were taken using a confocal laser scanning microscope at 488 nm, in duplicates A. BL21 pAC-p''lac''-''gfp'', positive control. B. BL21 pSB1C3-p''lac''-''gfp''. C. BL21, negative control.]] | [[File:BBa_K1804001_Fluorescence.png|600px|thumb|center|'''Figure 1:''' Representative images of bacterial cultures were taken using a confocal laser scanning microscope at 488 nm, in duplicates A. BL21 pAC-p''lac''-''gfp'', positive control. B. BL21 pSB1C3-p''lac''-''gfp''. C. BL21, negative control.]] |
Revision as of 15:02, 19 September 2015
GFP under the control of constitutive promoter lacI
Green fluorescence protein (GFP) from Aequeora victoria was placed under the control of the constitutive promoter lacI (plac).
plac promoter is constitutively active in E. coli BL21
The plac promoter is constitutively active in our strain of E. coli BL21. After construction of the pSB1C3-plac-gfp vector, the activity of the plac promoter was measured by quantification of fluorescence intensity. BL21 was used as the negative control and BL21 transformed with pAC-plac-gfp [[1]] as the positive control. As seen in Figures 1 and 2, GFP is expressed in both strains of BL21 containing the plac-gfp vectors but not in the negative control. When fluorescence data was quantified in arbitrary units (A.U.), the intensity of plac-gfp in pSB1C3 was lower relative to that of plac-gfp in the pAC vector (Figure 2). Nonetheless, our results indicated that plac is indeed able to drive gene expression under normal growth conditions. Supplementary Table 1 shows the descriptive statistics of the fluorescence intensities determined.
![](/wiki/images/5/5b/BBa_K1804001_FluorescenceIntensities.png)
Please refer to our Materials and Methods page [http://2015.igem.org/Team:SPSingapore/Anaerobic_Promoter] for details on fluorescence intensity quantification. Our experimental procedures can be found at our Notebook page [http://2015.igem.org/Team:SPSingapore/Notebook].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 734