Reporter

Part:BBa_K176016

Designed by: Chao Li, Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-06-30)
Revision as of 10:27, 21 October 2009 by Dqliu (Talk | contribs)

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pCon 0.01(J23103)->RBS+GFP+T

pCon 0.01(J23103)+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)

Model

To see the details of the model, click [http://2009.igem.org/Team:USTC/Modeling/Model-1 here] please.

data

J23103 + GFP

COLONY NUMBER T(min) OD600 FLU(arbitrary) ST.FLU(ustc_st.1) FLU/OD600(arbitrary) ST.FLU/OD600(ustc_st.1)
Ⅰ 0 0.343 0.218 0.871407443 0.635568513 2.540546481
Ⅰ 30 0.443 0.227 0.90738298 0.51241535 2.048268577
Ⅰ 60 0.519 0.316666667 1.265805919 0.61014772 2.438932406
Ⅰ 90 0.657 0.488666667 1.953338397 0.743784881 2.973117803
Ⅰ 120 0.691 0.577666667 2.309096481 0.835986493 3.341673634
Ⅰ 150 0.907 0.757333333 3.027274786 0.834987137 3.337678927
Ⅰ 180 1.366 1.305 5.216452812 0.95534407 3.818779511
Ⅱ 0 0.323 0.168333333 0.672875778 0.521155831 2.083206742
Ⅱ 30 0.418 0.245666667 0.981998907 0.587719298 2.349279683
Ⅱ 60 0.55 0.350666667 1.401713501 0.637575758 2.548570003
Ⅱ 90 0.621 0.412666667 1.649544976 0.664519592 2.656272103
Ⅱ 120 0.666 0.532 2.126553943 0.798798799 3.193023939
Ⅱ 150 0.888 0.721 2.882040213 0.811936937 3.24554078
Ⅱ 180 1.392 1.289333333 5.15382873 0.926245211 3.702463168 
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 0.01(J23103)->RBS+GFP+T