Difference between revisions of "Part:BBa K317040"

Latest revision as of 23:44, 27 October 2010

Pconst.(L)-RBS-LuxI-Double term-Plac-Lysis

This devicce includes BBa_K112808, the lysis device using three genes from enterobacteria phage T4: lysozyme, holin, and antiholin.

Evaluation of e BBa_K317040 for the characterization and application of BBa_K112808

We performed a experiment with this device in order to characterize BBa_K112808 activity.The experiment was performed from single colonies in a streaked LB agar plate.At first, we cultured the single colony having the above device in 100 mL LB for 12 hors at 37℃ with agitation.Then, the culture was dispensed into 3 mL scale and IPTG(f.c. 0.3 mM) was added to each dispensed culture to induce the expression of Lysis genes. The obtained culture was incubated for 20 hours at room temperature without agitation.After that, the OD660 of each culture(N=3) was measured.The result was shown below(Fig).

The result showed that the OD660 were different in presence of IPTG ( orange bar) or not (gray bar). OD660 of samples with the IPTG were half less than no additive samples. That indicates IPTG induced the expression of lysis genes and the E.coli in the culture were lysed.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 2850
Illegal NheI site found at 2873
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 2455
Illegal AgeI site found at 2525
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 3106

@@ Line 6: / Line 6: @@
 Evaluation of e BBa_K317040 for the characterization and application of BBa_K112808
-We performed a experiment with our parts above (Fig.3(B)) in order to characterize BBa_K112808 activity.The experiment was performed from single colonies in a streaked LB agar plate.At first, we cultured the single colony having the above device in 100 mL LB for 12 hors at 37℃ with agitation.Then, the culture was dispensed into 3 mL scale and IPTG(f.c. 0.3 mM) was added to each dispensed culture to induce the expression of Lysis genes. The obtained culture was incubated for 20 hours at room temperature without agitation.After that, the OD660 of each culture(N=3) was measured.The result was shown below(Fig).
+We performed a experiment with this device in order to characterize BBa_K112808 activity.The experiment was performed from single colonies in a streaked LB agar plate.At first, we cultured the single colony having the above device in 100 mL LB for 12 hors at 37℃ with agitation.Then, the culture was dispensed into 3 mL scale and IPTG(f.c. 0.3 mM) was added to each dispensed culture to induce the expression of Lysis genes. The obtained culture was incubated for 20 hours at room temperature without agitation.After that, the OD660 of each culture(N=3) was measured.The result was shown below(Fig).
 [[Image:IPTG lysis.jpg]]