Reporter

Part:BBa_K176015

Designed by: Chao Li, Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-06-30)
Revision as of 07:13, 20 October 2009 by Dqliu (Talk | contribs)

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)

data

J23109 + GFP

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


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