Difference between revisions of "Part:BBa K649104:Experience"
(→Applications of BBa_K649104) |
(→Applications of BBa_K649104) |
||
| Line 6: | Line 6: | ||
| − | Fluorescence intensity of our promoter lsrA-gfp(BBa_649104) was much higher than that of promoterless-gfp(negative control), showing that our new promoter lsrA(BBa_649100) works. On the other hand, fluorescence intensity of PlsrA(BBa_117002)-gfp was almost the same as a promoterless-gfp(negative control), showing that BBa_K117002 does not work properly. | + | Fluorescence intensity of our promoter lsrA-gfp(BBa_649104) was much higher than that of promoterless-gfp(negative control), showing that our new promoter lsrA(BBa_649100) works. On the other hand, fluorescence intensity of PlsrA(BBa_117002)-gfp was almost the same as a promoterless-gfp(negative control), showing that BBa_K117002 does not work properly.The difference between promoter lsrA(BBa_K117002) and promoter lsrA(BBa_649100) is whether promoter contains CRP binding site or not. Our promoter lsrA(BBa_649100) contains this site. According to Wang(2005) et al, cAMP-CRP directly binds to the upstream of promoter and stimulates expression of the lsr operon. |
| − | + | ||
Revision as of 10:08, 3 October 2011
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_K649104
Fluorescence intensity of our promoter lsrA-gfp(BBa_649104) was much higher than that of promoterless-gfp(negative control), showing that our new promoter lsrA(BBa_649100) works. On the other hand, fluorescence intensity of PlsrA(BBa_117002)-gfp was almost the same as a promoterless-gfp(negative control), showing that BBa_K117002 does not work properly.The difference between promoter lsrA(BBa_K117002) and promoter lsrA(BBa_649100) is whether promoter contains CRP binding site or not. Our promoter lsrA(BBa_649100) contains this site. According to Wang(2005) et al, cAMP-CRP directly binds to the upstream of promoter and stimulates expression of the lsr operon.
…TGTGAtctattcgTCGGA…
CRP recognition sites are shown in capital letter.BBa_K649100 contains this sites.
[sample]
pSB1A2 Ptet-gfp(JD22597)
pSB6A1 promoterless-gfp(JD22597)
pSB1A2 PlsrA-gfp(BBa_K649104)(JD22597)
pSB1A2 PlsrA-gfp(BBa_K11702-GFP)(JD22597)
[Method]
1.Overnight cultures of reporter strains grown at 37 °C in LB medium containing appropriate antibiotics were diluted 1:100 into 3 ml of LB medium and were incubated at 37 °C as fresh cultures.
2. After their OD590 reached 0.15, the fresh cultures were diluted 1:100.
3. After 4-hour incubation at 37 °C, 1 ml of each culture was moved to 1.6ml tube and its fluorescence intensity was measured with a flow cytometer
User Reviews
UNIQe88d0c5e15c9d9c1-partinfo-00000000-QINU UNIQe88d0c5e15c9d9c1-partinfo-00000001-QINU