Reporter

Part:BBa_K176013

Designed by: Chao Li, Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-06-30)
Revision as of 06:56, 20 October 2009 by Dqliu (Talk | contribs)

pCon 0.36(K176009)->RBS+GFP+T

pCon 0.36(K176009)+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.

data

J23107+GFP

COLONY NUMBER T(min) OD600 FLU(arbitrary) ST.FLU(ustc_st.1) FLU/OD600(arbitrary) ST.FLU/OD600(ustc_st.1)
Ⅰ 0 0.335 75.804 303.0099532 226.280597 904.5073231
Ⅰ 30 0.405 95.60666667 382.1667932 236.0658436 943.6217117
Ⅰ 60 0.485 119 475.67654 245.3608247 980.776371
Ⅰ 90 0.568 143.9033333 575.2221822 253.350939 1012.71511
Ⅰ 120 0.718 199.35 796.8581365 277.6462396 1109.830274
Ⅰ 180 0.984 266.53 1065.395531 270.8638211 1082.719036
Ⅱ 0 0.329 75.393 301.3670704 229.1580547 916.0093325
Ⅱ 30 0.404 101.95 407.5228844 252.3514851 1008.720011
Ⅱ 60 0.483 121.59 486.0294999 251.7391304 1006.272257
Ⅱ 90 0.565 147.0266667 587.7070259 260.2241888 1040.189426
Ⅱ 120 0.711 207.9833333 831.3680031 292.5222691 1169.293957
Ⅱ 180 0.97 273.8633333 1094.708931 282.3333333 1128.565909 
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 0.36(K176009)->RBS+GFP+T