Difference between revisions of "Part:BBa K4239008"
| Line 7: | Line 7: | ||
<partinfo>BBa_K4239008 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4239008 SequenceAndFeatures</partinfo> | ||
| + | <h2>Description</h2> | ||
| − | The <i>fiatlux operon</i> is composed of 6 genes: <i>fiatluxA, fiatluxB, fiatluxC, fiatluxD</i> and <i>fiatluxE</i>, that form an autonomous system producing bioluminescence thanks to the encoded luciferase enzymes. | + | <p>The <i>fiatlux operon</i> is composed of 6 genes: <i>fiatluxA, fiatluxB, fiatluxC, fiatluxD</i> and <i>fiatluxE</i>, that form an autonomous system producing bioluminescence thanks to the encoded luciferase enzymes.</p> |
| − | The genes <i>fiatluxA</i> and <i>fiatluxB</i> code each of them for a subunit of the luciferase protein. Both subunits need to be used together. Luciferase has as substrats FMNH2, O2 and Fatty aldehydes, and produces H20, Fatty Acids and FMN and emits luminescence. | + | <p>The genes <i>fiatluxA</i> and <i>fiatluxB</i> code each of them for a subunit of the luciferase protein. Both subunits need to be used together. Luciferase has as substrats FMNH2, O2 and Fatty aldehydes, and produces H20, Fatty Acids and FMN and emits luminescence.</p> |
| − | The genes <i>fiatluxC, fiatluxD</i> and <i>fiatluxE</i> code each of them for a subpart of a fatty acid reductase. They need to be used together to form a complex that recycles fatty acids to fatty aldehydes. Fatty aldehydes will be used as a substrate for the luciferase protein. | + | <p>The genes <i>fiatluxC, fiatluxD</i> and <i>fiatluxE</i> code each of them for a subpart of a fatty acid reductase. They need to be used together to form a complex that recycles fatty acids to fatty aldehydes. Fatty aldehydes will be used as a substrate for the luciferase protein.</p> |
| − | Both systemes (<i>fiatluxC/fiatluxD/fiatluxE</i> and <i>fiatluxA/fiatluxB</i>) are made to be used together, and are gathered in the <i>fiatluxCDABE</i> operon. | + | <p>Both systemes (<i>fiatluxC/fiatluxD/fiatluxE</i> and <i>fiatluxA/fiatluxB</i>) are made to be used together, and are gathered in the <i>fiatluxCDABE</i> operon.</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><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> |
| − | The <i>ilux</i> operon 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>The <i>ilux</i> operon 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> |
| + | <h2>References</h2> | ||
| + | |||
| + | <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> | ||
Revision as of 16:35, 9 October 2022
fiatlux genes with their promoter to emit luminescence
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 5342
Illegal BamHI site found at 50 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2993
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1366
Illegal SapI site found at 3512
Description
The fiatlux operon is composed of 6 genes: fiatluxA, fiatluxB, fiatluxC, fiatluxD and fiatluxE, that form an autonomous system producing bioluminescence thanks to the encoded luciferase enzymes.
The genes fiatluxA and fiatluxB code each of them for a subunit of the luciferase protein. Both subunits need to be used together. Luciferase has as substrats FMNH2, O2 and Fatty aldehydes, and produces H20, Fatty Acids and FMN and emits luminescence.
The genes fiatluxC, fiatluxD and fiatluxE code each of them for a subpart of a fatty acid reductase. They need to be used together to form a complex that recycles fatty acids to fatty aldehydes. Fatty aldehydes will be used as a substrate for the luciferase protein.
Both systemes (fiatluxC/fiatluxD/fiatluxE and fiatluxA/fiatluxB) are made to be used together, and are gathered in the fiatluxCDABE 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.
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.