Reporter

Part:BBa_K176014

Designed by: Chao Li, Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-06-30)
Revision as of 07:07, 20 October 2009 by Dqliu (Talk | contribs)

pCon 0.15(K176008)->RBS+GFP+T

pCon 0.15(K176008)+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.
K176011: J23100 + GFP ( I13504 )
K176012: J23101 + GFP ( I13504)
K176013: K176009( J23107) + GFP ( I13504)
K176014: K176008( J23115) + GFP ( I13504)
K176015: J23109 + GFP ( I13504)
K176016: J23103 + GFP ( I13504)

data

K176008+GFP

COLONY NUMBER T(min) OD600 FLU(arbitrary) ST.FLU(ustc_st.1) FLU/OD600(arbitrary) ST.FLU/OD600(ustc_st.1)
Ⅰ 0 0.344 6.204666667 24.80180144 18.03682171 72.09826
Ⅰ 30 0.451 9.088666667 36.32996229 20.15225425 80.55424012
Ⅰ 60 0.555 11.31433333 45.22657926 20.38618619 81.489332
Ⅰ 90 0.65 13.382 53.49162569 20.58769231 82.29480876
Ⅰ 120 0.685 15.55033333 62.15906517 22.70121655 90.74316083
Ⅰ 150 0.888 20.59866667 82.33867637 23.1966967 92.72373465
Ⅰ 180 1.281 29.948 119.7105968 23.37861046 93.45089523
Ⅱ 0 0.358 6.817666667 27.25213521 19.04376164 76.12328272
Ⅱ 30 0.456 9.824 39.26929688 21.54385965 86.11687912
Ⅱ 60 0.55 11.94633333 47.75286139 21.72060606 86.82338434
Ⅱ 90 0.647 15.013 60.01119239 23.20401855 92.75300215
Ⅱ 120 0.679 16.711 66.79857697 24.61119293 98.37787477
Ⅱ 150 0.866 22.18966667 88.69835179 25.62317167 102.423039
Ⅱ 180 1.247 31.647 126.5019787 25.37850842 101.445051 
For the details of the definition of ST.FLU and the unit ustc_st1, please click here.

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.


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Categories
Parameters
n/apCon 0.15(K176008)->RBS+GFP+T