Reporter

Part:BBa_K176015

Designed by: Chao Li, Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-06-30)

pCon 0.04(J23109)->RBS+GFP+T

pCon 0.04(J23109)+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

J23109 + GFP

COLONY NUMBER T(min) OD600 FLU(arbitrary) ST.FLU(ustc_st.1) FLU/OD600(arbitrary) ST.FLU/OD600(ustc_st.1)
0 0.383 0.355666667 1.421699911 0.928633594 3.712010211
30 0.495 0.509333333 2.035948888 1.028956229 4.113028057
60 0.592 0.614 2.454331055 1.037162162 4.145829485
90 0.703 0.786 3.141863533 1.118065434 4.469222664
120 0.746 0.989 3.953311748 1.325737265 5.299345507
150 0.958 1.309333333 5.233774367 1.366736256 5.463230028
180 1.338 2.187 8.742055402 1.634529148 6.533673694
0 0.294 0.229333333 0.916709971 0.780045351 3.118061124
30 0.399 0.344333333 1.376397383 0.86299081 3.449617502
60 0.491 0.427666667 1.709504204 0.871011541 3.481678623
90 0.581 0.572 2.286445217 0.984509466 3.93536182
120 0.625 0.729 2.914018467 1.1664 4.662429548
150 0.835 0.964333333 3.854712129 1.15489022 4.616421711
180 1.213 1.821666667 7.2817151 1.501786205 6.003062737
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.


[edit]
Categories
Parameters
n/apCon 0.04(J23109)->RBS+GFP+T