Reporter

Part:BBa_K176011

Designed by: Chao Li, Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-06-30)
Revision as of 07:08, 20 October 2009 by Dqliu (Talk | contribs)

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)

data

J23100+GFP

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


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Categories
Parameters
n/apCon 1.00(J23100)->RBS+GFP+T