Reporter

Part:BBa_K176012

Designed by: Chao Li, Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-06-30)
Revision as of 06:45, 20 October 2009 by Dqliu (Talk | contribs)

pCon 0.70(J23101)->RBS+GFP+T

pCon 0.70(J23101)+I13504

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
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 705

MEASUREMENT

We measured five constitutive promoter family member ( J23100, J23101, J23103, J23107, J23109, J23115, )strength by ligated each with a 
GFP gene ( I13504) , and then moniter the OD600 and GFP expression.

data

J23101+GFP

COLONY NUMBER T(min) OD600 FLU(arbitrary) ST.FLU(ustc_st.1) FLU/OD600(arbitrary) ST.FLU/OD600(ustc_st.1)
Ⅰ 0 0.117666667 142.91 571.2515489 1214.532578 4854.829028
Ⅰ 30 0.119666667 152.6266667 610.0918042 1275.431755 5098.260203
Ⅰ 60 0.143666667 181.65 726.1062478 1264.385151 5054.103813
Ⅰ 90 0.174666667 206.08 823.7598433 1179.847328 4716.182309
Ⅰ 120 0.198 220.6666667 882.0668612 1114.478114 4454.883137
Ⅰ 150 0.245666667 242.8866667 970.8864639 988.6838535 3952.048021
Ⅰ 180 0.392 276.32 1104.52892 704.8979592 2817.675817
Ⅱ 0 0.083 115.0533333 459.9006009 1386.184739 5540.971095
Ⅱ 30 0.098333333 126.3533333 505.0698858 1284.949153 5136.303924
Ⅱ 60 0.112 144.0466667 575.795126 1286.130952 5141.027911
Ⅱ 90 0.129666667 188.7433333 754.4603003 1455.604113 5818.4599
Ⅱ 120 0.14 174.4166667 697.1925757 1245.833333 4979.946969
Ⅱ 150 0.169333333 184.28 736.619099 1088.267717 4350.112789
Ⅱ 180 0.251333333 232.6733333 930.0608919 925.7559682 3700.507528

plot

Promoters.jpg

protocol

1. Streak a plate of the strain which contain one of the parts listed in pSB1A3 .

2. Inoculate two 3ml cultures of supplemented M9 Medium and antibiotic( Ampicillin 0.1mg/ml) with single colony from the plate.

3. Cultures were grown in test tubes（BIO BASIC INC.12ml Polypropylene Round-bottom Culture Tubes With Graduations And Dual Cap Cat.No:TD444） for 16hrs at 37℃ with shaking at 200rpm.

4. Cultures were diluted 1:100 into 3ml fresh medium and grown for 3hrs.

5. Measure the fluorescence(SHIMDZU SPECTROFLUOROPHOTOMETER RF-5301PC, 250ul quartz cell path length 10mm,501 nm excitation,514 nm emission,1.5nm slit width) and absorbance (HITACHI UV-VIS spectrophotometer U-2810 ,200ul quartz cell,path length 10mm,600nm,1.5 nm slit width) every 30 minutes in the next 4hrs.


[edit]
Categories
Parameters
n/apCon 0.70(J23101)->RBS+GFP+T