Difference between revisions of "Part:BBa K176015"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K176015 short</partinfo>
 
<partinfo>BBa_K176015 short</partinfo>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K176015 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K176015 SequenceAndFeatures</partinfo>
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=='''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 )
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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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J23109 + 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||0.383||0.355666667||1.421699911||0.928633594||3.712010211
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|-
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| Ⅰ||30||0.495||0.509333333||2.035948888||1.028956229||4.113028057
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|-
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| Ⅰ||60||0.592||0.614||2.454331055||1.037162162||4.145829485
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|-
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| Ⅰ||90||0.703||0.786||3.141863533||1.118065434||4.469222664
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|-
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| Ⅰ||120||0.746||0.989||3.953311748||1.325737265||5.299345507
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|-
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| Ⅰ||150||0.958||1.309333333||5.233774367||1.366736256||5.463230028
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|-
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| Ⅰ||180||1.338||2.187||8.742055402||1.634529148||6.533673694
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|-
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| Ⅱ||0||0.294||0.229333333||0.916709971||0.780045351||3.118061124
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|-
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| Ⅱ||30||0.399||0.344333333||1.376397383||0.86299081||3.449617502
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|-
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| Ⅱ||60||0.491||0.427666667||1.709504204||0.871011541||3.481678623
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|-
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| Ⅱ||90||0.581||0.572||2.286445217||0.984509466||3.93536182
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|-
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| Ⅱ||120||0.625||0.729||2.914018467||1.1664||4.662429548
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|-
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| Ⅱ||150||0.835||0.964333333||3.854712129||1.15489022||4.616421711
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|-
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| Ⅱ||180||1.213||1.821666667||7.2817151||1.501786205||6.003062737
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|}
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{{USTC//promoters}}
  
  

Revision as of 07:13, 20 October 2009

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.