Reporter

Part:BBa_K176011

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

pCon 1.00(J23100)->RBS+GFP+T

pCon 1.00(J23100)+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, please click here

data

J23100+GFP

COLONY NUMBER T(min) OD600 FLU(arbitrary) ST.FLU(ustc_st.1) FLU/OD600(arbitrary) ST.FLU/OD600(ustc_st.1)
0 0.071 132.07 527.9210137 1860.140845 7435.507235
30 0.076 144.49 577.5672543 1901.184211 7599.569135
60 0.101 157.6633333 630.2247805 1561.023102 6239.849312
90 0.093 160.8866667 643.1093523 1729.964158 6915.154326
120 0.116 176.33 704.8407083 1520.086207 6076.213003
150 0.122666667 185.0633333 739.7503031 1508.668478 6030.573123
180 0.153 197.8966667 791.0487535 1293.442266 5170.253291
0 0.07 144.1 576.0083143 2058.571429 8228.690205
30 0.075333333 136.1066667 544.0567081 1806.725664 7221.991701
60 0.084333333 144.92 579.2860855 1718.418972 6869.004966
90 0.092 157.4133333 629.22546 1711.014493 6839.407174
120 0.098 164.73 658.4722389 1680.918367 6719.104478
150 0.117 179.78 718.6313307 1536.581197 6142.148126
180 0.135666667 194.6566667 778.0975603 1434.815725 5735.362853
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 1.00(J23100)->RBS+GFP+T