Difference between revisions of "Part:BBa K176016"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K176016 short</partinfo>
 
<partinfo>BBa_K176016 short</partinfo>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K176016 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K176016 SequenceAndFeatures</partinfo>
 +
=='''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.
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K176011: J23100 + GFP ( I13504 )
 +
K176012: J23101 + GFP ( I13504)
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K176013: K176009( J23107) + GFP ( I13504)
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K176014: K176008( J23115) + GFP ( I13504)
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K176015: J23109 + GFP ( I13504)
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K176016: J23103 + GFP ( I13504)
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===data===
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J23103 + GFP
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{| {{table}}
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{|class="sortable" style="border:3px solid black; width: 70%" align="center"
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| align="center" style="background:#f0f0f0;"|'''COLONY NUMBER'''
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| align="center" style="background:#f0f0f0;"|'''T(min)'''
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| align="center" style="background:#f0f0f0;"|'''OD600'''
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| align="center" style="background:#f0f0f0;"|'''FLU(arbitrary)'''
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| align="center" style="background:#f0f0f0;"|'''ST.FLU(ustc_st.1)'''
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| align="center" style="background:#f0f0f0;"|'''FLU/OD600(arbitrary)'''
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| align="center" style="background:#f0f0f0;"|'''ST.FLU/OD600(ustc_st.1)'''
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|-
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| Ⅰ||0||0.343||0.218||0.871407443||0.635568513||2.540546481
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|-
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| Ⅰ||30||0.443||0.227||0.90738298||0.51241535||2.048268577
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|-
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| Ⅰ||60||0.519||0.316666667||1.265805919||0.61014772||2.438932406
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|-
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| Ⅰ||90||0.657||0.488666667||1.953338397||0.743784881||2.973117803
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|-
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| Ⅰ||120||0.691||0.577666667||2.309096481||0.835986493||3.341673634
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|-
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| Ⅰ||150||0.907||0.757333333||3.027274786||0.834987137||3.337678927
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|-
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| Ⅰ||180||1.366||1.305||5.216452812||0.95534407||3.818779511
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|-
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| Ⅱ||0||0.323||0.168333333||0.672875778||0.521155831||2.083206742
 +
|-
 +
| Ⅱ||30||0.418||0.245666667||0.981998907||0.587719298||2.349279683
 +
|-
 +
| Ⅱ||60||0.55||0.350666667||1.401713501||0.637575758||2.548570003
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|-
 +
| Ⅱ||90||0.621||0.412666667||1.649544976||0.664519592||2.656272103
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|-
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| Ⅱ||120||0.666||0.532||2.126553943||0.798798799||3.193023939
 +
|-
 +
| Ⅱ||150||0.888||0.721||2.882040213||0.811936937||3.24554078
 +
|-
 +
| Ⅱ||180||1.392||1.289333333||5.15382873||0.926245211||3.702463168
 +
|}
 +
{{USTC//promoters}}
  
  

Revision as of 07:18, 20 October 2009

pCon 0.01(J23103)->RBS+GFP+T

pCon 0.01(J23103)+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

J23103 + GFP

COLONY NUMBER T(min) OD600 FLU(arbitrary) ST.FLU(ustc_st.1) FLU/OD600(arbitrary) ST.FLU/OD600(ustc_st.1)
Ⅰ 0 0.343 0.218 0.871407443 0.635568513 2.540546481
Ⅰ 30 0.443 0.227 0.90738298 0.51241535 2.048268577
Ⅰ 60 0.519 0.316666667 1.265805919 0.61014772 2.438932406
Ⅰ 90 0.657 0.488666667 1.953338397 0.743784881 2.973117803
Ⅰ 120 0.691 0.577666667 2.309096481 0.835986493 3.341673634
Ⅰ 150 0.907 0.757333333 3.027274786 0.834987137 3.337678927
Ⅰ 180 1.366 1.305 5.216452812 0.95534407 3.818779511
Ⅱ 0 0.323 0.168333333 0.672875778 0.521155831 2.083206742
Ⅱ 30 0.418 0.245666667 0.981998907 0.587719298 2.349279683
Ⅱ 60 0.55 0.350666667 1.401713501 0.637575758 2.548570003
Ⅱ 90 0.621 0.412666667 1.649544976 0.664519592 2.656272103
Ⅱ 120 0.666 0.532 2.126553943 0.798798799 3.193023939
Ⅱ 150 0.888 0.721 2.882040213 0.811936937 3.24554078
Ⅱ 180 1.392 1.289333333 5.15382873 0.926245211 3.702463168 
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.