Difference between revisions of "Part:BBa K143082"

Revision as of 22:08, 29 October 2008

Pveg-spoVG RFP expression construct

RFP gene under expresson of the Pveg promoter and spoVG RBS of B. subtilis, with a chloramphenicol resistance marker for ease of selection.

Also contains the 5' and 3' amyE integration sequences to allow integration into B. subtilis at the amyE locus.

This device was used by the Imperial iGEM 2008 team to characterise the Pveg promoter and spoVG RBS (Combined part BBa_K143053) as part of the Biofabricator subtilis project.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 2136
Illegal AgeI site found at 2248
1000
COMPATIBLE WITH RFC[1000]

Part Characterisation

This test construct was used for the characterise the expression output of the promoter pVeg combined with the ribosome binding site spoVG. The characterisation was performed using a plate reader to measure fluorescence and O.D.₆₀₀. The O.D.₆₀₀ data was converted to cell number (colony forming units) using the calibration curve O.D.₆₀₀ vs cell number. Cell number was then used to normalise the fluorescence to give the final normalised units of fluorescence per cell (colony forming unit). The graph below shows the results of a B.subtilis culture containing this test construct and a culture of B.subtilis containing the test construct BBa_K143078 as a negative control. It is clear that B.subtilis containing this test construct is expressing mRFP. To learn more about the characterisation please [http://2008.igem.org/Team:Imperial_College/Biobricks click this link.]

@@ Line 28: / Line 28: @@
 <span class='h3bb'><big>'''Part Characterisation'''</big></span>
+[[Image:B.subtilisRFP.PNG|400px|center]]
+This test construct was used for the characterise the expression output of the promoter pVeg combined with the ribosome binding site spoVG. The characterisation was performed using a plate reader to measure fluorescence and O.D.<sub>600</sub>. The O.D.<sub>600</sub> data was converted to cell number (colony forming units) using the calibration curve O.D.<sub>600</sub> vs cell number. Cell number was then used to normalise the fluorescence to give the final normalised units of fluorescence per cell (colony forming unit). The graph below shows the results of a ''B.subtilis'' culture containing this test construct and a culture of ''B.subtilis'' containing the test construct [https://parts.igem.org/wiki/index.php?title=Part:BBa_K143079 BBa_K143078] as a negative control. It is clear that ''B.subtilis'' containing this test construct is expressing mRFP. To learn more about the characterisation please [http://2008.igem.org/Team:Imperial_College/Biobricks click this link.]