Difference between revisions of "Part:BBa Q04510"

 
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_Q04510 short</partinfo>
 
<partinfo>BBa_Q04510 short</partinfo>
  
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Lambda cI QPI w/ strong RBS
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Lambda cI [[QPI]] w/ strong RBS
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===Usage and Biology===
 
===Usage and Biology===
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Preliminary data indicates that this inverter functions well. [jb, 5/24/04]
 
Preliminary data indicates that this inverter functions well. [jb, 5/24/04]
  
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<partinfo>BBa_Q04510 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_Q04510 SequenceAndFeatures</partinfo>
  
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<!-- Uncomment this to enable Functional Parameter display
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_Q04510 parameters</partinfo>
 
<partinfo>BBa_Q04510 parameters</partinfo>
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<!-- -->
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==USTC_2009's MEASUREMENT==
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[https://parts.igem.org/Part:BBa_K176112 K176112]
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[https://parts.igem.org/Part:BBa_K176113 K176113]
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[https://parts.igem.org/Part:BBa_K176114 K176114]
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=ECUST_China 2019 characterization=
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In ECUST_China 2019 characterization, we added the Plac before the CⅠgene and used mRFP as the reporter gene to measure the effectiveness of this part.
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In the absence of IPTG, CⅠP was constitutively expressed so that the RFP fluorescence could be observed. However, as the concentration of IPTG increasing, Plac was activated to express CⅠprotein which inhibited the transcription of CⅠP ，so the fluorescence of mRFP decreased.
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=Contribution=
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'''Group''': [https://2020.igem.org/Team/XMU-China iGEM Team XMU-China 2020]
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'''Author''': Shi Zhang
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'''Summary''': fluorescence / OD<sub>600</sub>
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===Quantitative Characterization from iGEM20-XMU-China===
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Inverter is mainly composed of cI repressor from ''E. coli'' phage lambda and pR promoter which is inhibited by cI repressor. It can reverse the inducer action of the upstream  promoter. It was registered in 2003.
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<html>
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    <figure>
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        <img src="https://2020.igem.org/wiki/images/c/c7/T--XMU-China--XMU-China_2020_inverter.png" width="40%" style="float:center">
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        <figcaption>
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        <p style="font-size:1rem">
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        </p>
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        </figcaption>
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    </figure>
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</html>
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:'''Fig.1''' Genetic circuits of Inverter.
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We constructed “P<sub>Bad/araC</sub>-RBS-EYFP-pSB1C3”(<partinfo>BBa_K3332082</partinfo>) and “P<sub>Bad/araC</sub>-Inverter-RBS-EYFP-pSB1C3”(<partinfo>BBa_K3332079</partinfo>) in ''E.coli'' BL21（DE3）to characterize its function.
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;For the latter circuit, cI repressor will not express in the absence of arabinose so that the fluorescence of EYFP can be observed. At the certain concentration of arabinose, PBad/araC can be activated to express cI repressor to inhibit the pR promoter，so the fluorescence of EYFP decreases.
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The fluorescence/OD<sub>600</sub> values of these two circuits were measured with or without arabinose and compared with “P<sub>Bad/araC</sub>-pSB1C3” in ''E.coli'' BL21(DE3). We discovered that the downstream gene of proBAD/araC can be expressed in the presence of arabinose and decrease expression without arabinose. When the inverter is added to the circuit, the effect is reversed. The result proves that inverter can function correctly and reverse the effect of P<sub>Bad/araC</sub>. 
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<html>
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    <figure>
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        <img src="https://2020.igem.org/wiki/images/3/3d/T--XMU-China--XMU-China_2020-BY_BNY_fluorescence.png" width="60%" style="float:center">
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        <figcaption>
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        <p style="font-size:1rem">
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        </p>
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        </figcaption>
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    </figure>
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</html>
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:'''Fig.2''' The results of Fluorescence Intensity/OD.Yellow represents P<sub>Bad/araC</sub>, green represents P<sub>Bad/araC</sub>-EYFP(<partinfo>BBa_K3332082</partinfo>) , and purple represents P<sub>Bad/araC</sub> -Inverter-EYFP(<partinfo>BBa_K3332079</partinfo>) .

Latest revision as of 22:03, 27 October 2020

QPI (B0034.C0051.B0015.R0051)

Lambda cI QPI w/ strong RBS


Usage and Biology

Preliminary data indicates that this inverter functions well. [jb, 5/24/04]

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


USTC_2009's MEASUREMENT

K176112

K176113

K176114

ECUST_China 2019 characterization

In ECUST_China 2019 characterization, we added the Plac before the CⅠgene and used mRFP as the reporter gene to measure the effectiveness of this part. In the absence of IPTG, CⅠP was constitutively expressed so that the RFP fluorescence could be observed. However, as the concentration of IPTG increasing, Plac was activated to express CⅠprotein which inhibited the transcription of CⅠP ，so the fluorescence of mRFP decreased.


Contribution

Group: iGEM Team XMU-China 2020

Author: Shi Zhang

Summary: fluorescence / OD600

Quantitative Characterization from iGEM20-XMU-China

        Inverter is mainly composed of cI repressor from E. coli phage lambda and pR promoter which is inhibited by cI repressor. It can reverse the inducer action of the upstream promoter. It was registered in 2003.

Fig.1 Genetic circuits of Inverter.


        We constructed “PBad/araC-RBS-EYFP-pSB1C3”(BBa_K3332082) and “PBad/araC-Inverter-RBS-EYFP-pSB1C3”(BBa_K3332079) in E.coli BL21（DE3）to characterize its function.

        For the latter circuit, cI repressor will not express in the absence of arabinose so that the fluorescence of EYFP can be observed. At the certain concentration of arabinose, PBad/araC can be activated to express cI repressor to inhibit the pR promoter，so the fluorescence of EYFP decreases.

        The fluorescence/OD600 values of these two circuits were measured with or without arabinose and compared with “PBad/araC-pSB1C3” in E.coli BL21(DE3). We discovered that the downstream gene of proBAD/araC can be expressed in the presence of arabinose and decrease expression without arabinose. When the inverter is added to the circuit, the effect is reversed. The result proves that inverter can function correctly and reverse the effect of PBad/araC.

Fig.2 The results of Fluorescence Intensity/OD.Yellow represents PBad/araC, green represents PBad/araC-EYFP(BBa_K3332082) , and purple represents PBad/araC -Inverter-EYFP(BBa_K3332079) .