Composite

Part:BBa_M36938:Experience

Designed by: Emily Kelly   Group: Stanford BIOE44 - S11   (2014-10-23)
Revision as of 06:54, 4 December 2014 by CME2 (Talk | contribs)

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Applications of BBa_M36938

Stanford Location:

Box Name: BIOE44 S14

Glycerol Freezer Stocks:

Barcode# :133011796, Plasmid Name: CME LCPUFA_Dasher, Resistance Type: amp, DNA 2.0 Gene #:193425, Organism: E. coli, Device Type: LCPUFA Actuator.

Barcode# : 133024687, Plasmid Name: CME LCPUFA_Dasher, Resistance Type: amp, DNA 2.0 Gene #:193425, Organism: E. coli, Device Type: LCPUFA Actuator.

Barcode# : 133010087, Plasmid Name: CME LCPUFA_Dasher, Resistance Type: amp, DNA 2.0 Gene #:193425, Organism: E. coli, Device Type: LCPUFA Actuator.

Barcode# : 133011361, Plasmid Name: CME LCPUFA_Dasher, Resistance Type: amp, DNA 2.0 Gene #:193425, Organism: E. coli, Device Type: LCPUFA Actuator.

Performance Data:

Note: The entries in the same numerical position in each list are data for one sample.

Protein Presence Test: GFP Fluorescence Reading

Condition GFP GFP GFP GFP Control Control Control Control Blank

OD600 1.44 1.377 1.422 1.377 1.432 1.448 1.45 1.451 0.037

GFP AFU 3537 4250 3725 3604 829 966 918 853 658

Standardized OD600 1.403 1.34 1.385 1.34 1.395 1.411 1.413 1.414 0

Standardized GFP AFU 2852 3565 3040 2919 144 281 233 168 0

AFU/OD600 2032.78689 2660.44776 2194.94585 2178.35821 103.225806 199.149539 164.897381 118.811881

GFP Average: 2266.63468 Control Average: 146.521152

p = 0.00036738 ---> p < 0.001


GFPchart.gif


GFP.jpg


Conclusion: The target protein (enoyl-[acyl-carrier-protein] reductase) is synthesized within the E. coli cells. This result is statistically significant.


Protein Functionality Test: pH Reading

Functional (enoyl-[acyl-carrier-protein] reductase) is expected to partake in a naturally occurring reaction within the E. coli cell. This reaction consumes hydrogen ions. Thus a functional protein is expected to increase the overall pH of the cell.

Note: pH Readings were not taken for this composite part but for its sister part BBa_M36939 (lacks Comet GFP). We suspect that similar results would be obtained for this construct (containing Comet GFP). This data is included simply to provide a full analysis of the mechanism and its potential.

Standard(no GFP) pH Readings: 6.91 6.64 6.49 6.42 6.64 6.45 6.34 6.4

Control pH Readings:

6.56 6.49 6.42 6.36 6.25 6.26 6.29 6.34

Standard Average pH: 6.53625

Control Average pH: 6.37125

p=0.011066469 ---> p < 0.05

Conclusion: Production of the target protein (enoyl-[acyl-carrier-protein] reductase) causes an increase in pH, as is to be expected. Therefore we conclude that the target protein is functional. This is statistically significant.

User Reviews

UNIQ7badff076a486be3-partinfo-00000000-QINU UNIQ7badff076a486be3-partinfo-00000001-QINU