Difference between revisions of "Part:BBa K4239004"
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<partinfo>BBa_K4239004 short</partinfo>
 
<partinfo>BBa_K4239004 short</partinfo>
  
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fiatluxB is made to be used with fiatluxA. It codes for a subpart of the luciferase protein. With the subpart coding from fiatluxA, they form the luciferase protein.
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<html>
  
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Luciferase has as substrats FMNH2, O2 and Fatty aldehydes, and produces H20, Fatty Acids and FMN and emits luminescence.
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<br>
  
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The systeme fiatluxA/fiatluxB is made to be used with fiatluxC, fiatluxC and fiatluxE, gathered in the fiatluxCDABE operon.
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<h2>Description</h2>
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<p><i>fiatluxB</i> is made to be used with
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<i>fiatluxA</i> <a href="https://parts.igem.org/Part:BBa_K239003" class="pr-0" target="_blank">(BBa_K239003)</a>.
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It codes for a subpart of the luciferase protein. With the subpart coding from <i>fiatluxA</i>, they form the luciferase protein.</p>
 +
 
 +
<p>Luciferase has as substrats FMNH2, O2 and Fatty aldehydes, and produces H20, Fatty Acids and FMN and emits luminescence.</p>
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 +
<p>The systeme <i>fiatluxA/fiatluxB</i> is made to be used with  
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<i>fiatluxC</i> <a href="https://parts.igem.org/Part:BBa_K239001" class="pr-0" target="_blank">(BBa_K239001)</a>,  
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<i>fiatluxD</i> <a href="https://parts.igem.org/Part:BBa_K239002" class="pr-0" target="_blank">(BBa_K239002)</a>
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and <i>fiatluxE</i> <a href="https://parts.igem.org/Part:BBa_K239005" class="pr-0" target="_blank">(BBa_K239005)</a>, gathered in the <i>fiatluxCDABE</i> <a href="https://parts.igem.org/Part:BBa_K239006" class="pr-0" target="_blank">(BBa_K239006)</a>
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operon.</p>
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<p><i>Fiatlux</i> genes come from <i>ilux</i> genes (C, D, A, B, E). They were modified to remove every Igem restriction site (EcoR1, Xba1, Spe1 and Pst1) included in genes. They were also adapted to include the biobrick format.</p>
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<p>The <i>ilux operon</i> was born from a mutated natural luminescence operon present in the bacteria P.luminescens: the <i>lux</i> operon. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018 (Gregor et al. 2018). The aim was to create a system of genes that produced more light.</p>
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<br>
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</html>
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<h2>Sequence and Features</h2>
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<partinfo>BBa_K4239004 SequenceAndFeatures</partinfo>
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<html>
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<br>
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<h2>Construction</h2>
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<p>The <i>ilux</i> operon was available in a pGEX plasmid. <i>fiatluxA, fiatluxB</i> and <i>fiatluxE</i> were directly constructed together in <i>fiatluxABE</i>. Igem restriction sites were successfully removed in the <i>iluxABE</i> genes by following these steps: DNA extraction, PCR directed mutagenesis, agarose gel analysis with green gel, and gel purification. An overlap PCR was performed to reconstitute <i>iluxABE</i> fragments which had been cut by the restriction enzymes. The part is now called <i>fiatluxABE</i> This part was then cloned and transformed in a pSB1C3 (already in iGEM biobrick format) and pBAD18 (high-copy vector with an arabinose inducible promoter) plasmids in E.coli DH5α. More details about the construction are on the following page
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<i>fiatluxABE</i> <a href="https://parts.igem.org/Part:BBa_K239007" class="pr-0" target="_blank">(BBa_K239007)</a>.
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<br>
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<br>
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<h2>References</h2>
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<p><i> Gregor C, Gwosch KC, Sahl SJ, Hell SW. Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging. Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):962-967. doi: 10.1073/pnas.1715946115. Epub 2018 Jan 16. PMID: 29339494; PMCID: PMC5798359.</i> </p>
  
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Fiatlux genes come from ilux genes (C, D, A, B, E). They were modified to remove every Igem restriction site (EcoR1, Xba1, Spe1 and Pst1) included in genes. They were also adapted to include the biobrick format.
 
  
−
The ilux operon was born from a mutated natural luminescence operon present in the bacteria P.luminescens: the lux operon. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018 (Gregor et al. 2018). The aim was to create a system of genes that produced more light.
 
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4239004 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K4239004 parameters</partinfo>
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Revision as of 07:38, 10 October 2022


Enhanced luciferase substrate forming unit fiatluxB


Description

fiatluxB is made to be used with fiatluxA (BBa_K239003). It codes for a subpart of the luciferase protein. With the subpart coding from fiatluxA, they form the luciferase protein.

Luciferase has as substrats FMNH2, O2 and Fatty aldehydes, and produces H20, Fatty Acids and FMN and emits luminescence.

The systeme fiatluxA/fiatluxB is made to be used with fiatluxC (BBa_K239001), fiatluxD (BBa_K239002) and fiatluxE (BBa_K239005), gathered in the fiatluxCDABE (BBa_K239006) operon.

Fiatlux genes come from ilux genes (C, D, A, B, E). They were modified to remove every Igem restriction site (EcoR1, Xba1, Spe1 and Pst1) included in genes. They were also adapted to include the biobrick format.

The ilux operon was born from a mutated natural luminescence operon present in the bacteria P.luminescens: the lux operon. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018 (Gregor et al. 2018). The aim was to create a system of genes that produced more light.


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]


Construction

The ilux operon was available in a pGEX plasmid. fiatluxA, fiatluxB and fiatluxE were directly constructed together in fiatluxABE. Igem restriction sites were successfully removed in the iluxABE genes by following these steps: DNA extraction, PCR directed mutagenesis, agarose gel analysis with green gel, and gel purification. An overlap PCR was performed to reconstitute iluxABE fragments which had been cut by the restriction enzymes. The part is now called fiatluxABE This part was then cloned and transformed in a pSB1C3 (already in iGEM biobrick format) and pBAD18 (high-copy vector with an arabinose inducible promoter) plasmids in E.coli DH5α. More details about the construction are on the following page fiatluxABE (BBa_K239007).

References

Gregor C, Gwosch KC, Sahl SJ, Hell SW. Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging. Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):962-967. doi: 10.1073/pnas.1715946115. Epub 2018 Jan 16. PMID: 29339494; PMCID: PMC5798359.

Sequence and Features