Difference between revisions of "Part:BBa K112019"

Latest revision as of 08:28, 6 November 2008

Lambda phage lysis device

Lysozyme and holin production are driven by pBad, and antiholin is driven by PconC5.

Characterization

For more info, visit [http://2008.igem.org/Team:UC_Berkeley/LysisDevice UC Berkeley iGEM08 Wiki!!]

K112019 (Lambda Lysis Device) was inserted in the vector K112950 and K112809 was insertedin pSB1A2. They were introduced into the MC1061 strain by transformation, and then picked 5 colonies for each device. We grew these cultures to saturation at 37 degrees Celsius in LB media, and then split into eight 1 mL aliquots. A range of concentrations of arabinose was added to these aliquots, with a starting concentration of 1.3E-3 M and the next 6 samples recieving a four-fold dilution of the previous sample, and an equal volume of water added to the last aliquot. The cultures were then incubated at 37 degrees again for 3.5 hours, and the absorbance at 600nm was measured with a Tecan Xfluor4 Safire2 in a Corning Inc. Costar 3603 plate. The data plotted on a log scale is shown below.

Data from adding arabinose at midlog. Concentrations used at each data point from left to right are 0, 2.44E-07, 9.77E-07, 3.91E-06, 1.56E-05, 6.25E-05, 2.50E-04, and 1.00E-03 M

A parallel experiment was performed by taking the same saturated culture before induction with arabinose, diluting 100-fold, growing to mid-log(starting OD .22), before inducing with arabinose as above. The data plotted on a log scale is shown below.

Data from adding arabinose at midlog. Concentrations used at each data point from left to right are 0, 2.44E-07, 9.77E-07, 3.91E-06, 1.56E-05, 6.25E-05, 2.50E-04, and 1.00E-03 M

This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at:
[http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 33
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 33
Illegal NheI site found at 1184
Illegal NheI site found at 2230
Illegal NheI site found at 2253
21
INCOMPATIBLE WITH RFC[21]
Illegal prefix found in sequence at 33
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 33
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 33
Illegal NgoMIV site found at 2616
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 1050

@@ Line 5: / Line 5: @@
 ==Characterization==
-We transformed this device in the vector K112950 into the MC1061 strain, and then picked 5 colonies for each device. We grew these cultures to saturation at 37 degrees Celsius in LB media, and then split into eight 1 mL aliquots. A range of concentrations of arabinose was added to these aliquots, with a starting concentration of 1.3E-3 M and the next 6 samples recieving a four-fold dilution of the previous sample, and an equal volume of water added to the last aliquot. The cultures were then incubated at 37 degrees again for 3.5 hours, and the absorbance at 600nm was measured with a Tecan Xfluor4 Safire2 in a Corning Inc. Costar 3603 plate. The data plotted on a log scale is shown below.
+'''For more info, visit [http://2008.igem.org/Team:UC_Berkeley/LysisDevice UC Berkeley iGEM08 Wiki!!]'''
+K112019 (Lambda Lysis Device) was inserted in the vector K112950 and K112809 was insertedin pSB1A2. They were introduced into the MC1061 strain by transformation, and then picked 5 colonies for each device. We grew these cultures to saturation at 37 degrees Celsius in LB media, and then split into eight 1 mL aliquots. A range of concentrations of arabinose was added to these aliquots, with a starting concentration of 1.3E-3 M and the next 6 samples recieving a four-fold dilution of the previous sample, and an equal volume of water added to the last aliquot. The cultures were then incubated at 37 degrees again for 3.5 hours, and the absorbance at 600nm was measured with a Tecan Xfluor4 Safire2 in a Corning Inc. Costar 3603 plate. The data plotted on a log scale is shown below.