Difference between revisions of "Part:BBa K4179020"
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<partinfo>BBa_K4179020 short</partinfo> | <partinfo>BBa_K4179020 short</partinfo> | ||
| − | The composite part comprises a Blasticidin S deaminase[https://parts.igem.org/Part:BBa_K1943015 (BBa_K1943015)], which is a Blasticidin resistance gene with the Kozak sequence. Downstream to this under the CMV[https://parts.igem.org/Part:BBa_I712004 (BBa_I712004)] which is a constitutive promoter, an SV40PolyA terminator[https://parts.igem.org/Part:BBa_K4235020 (BBa_K4235020)] is cloned. Together in this composite, a cTNT promoter[https://parts.igem.org/Part:BBa_K4179021 (BBa_K4179021)] is found, downstream to the 8XGTIIC motifs[https://parts.igem.org/Part:BBa_K4179017 (BBa_K4179017)] followed by the luciferase gene[https://parts.igem.org/Part:BBa_I712019 (BBa_I712019)] and the low most part SV40 Poly(A)[https://parts.igem.org/Part:BBa_K4235020 (BBa_K4235020)]. | + | The composite part comprises a Blasticidin S deaminase [https://parts.igem.org/Part:BBa_K1943015 (BBa_K1943015)], which is a Blasticidin resistance gene with the Kozak sequence. Downstream to this under the CMV [https://parts.igem.org/Part:BBa_I712004 (BBa_I712004)] which is a constitutive promoter, an SV40PolyA terminator [https://parts.igem.org/Part:BBa_K4235020 (BBa_K4235020)] is cloned. Together in this composite, a cTNT promoter [https://parts.igem.org/Part:BBa_K4179021 (BBa_K4179021)] is found, downstream to the 8XGTIIC motifs [https://parts.igem.org/Part:BBa_K4179017 (BBa_K4179017)] followed by the luciferase gene [https://parts.igem.org/Part:BBa_I712019 (BBa_I712019)] and the low most part SV40 Poly(A)[https://parts.igem.org/Part:BBa_K4235020 (BBa_K4235020)]. |
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===Usage and Biology=== | ===Usage and Biology=== | ||
The team of Technion 2022 used this composite as a part of their cell line measurement system called Oracell. This biological system quantifies their manufactured metabolite-decursin by utilizing its effect on the Hippo pathway which is a signaling pathway that regulated proliferation and apoptosis in mammalian cells [1]. TEAD transcriptional factors bind to the 8XGTIIC motifs and regulate the expression of the luciferase gene which is cloned downstream to these motifs. Luciferase is used as a reporter gene that emits luminescence. Thus, the quantity of this metabolite is measured according to the luminescence values, by using a calibration curve. This composite is transfected to CHO (Chinese Hamster Ovarian) cells by stable integration. The role of the Blasticidin S deaminase here is to accomplish a successful selection for positive cells after transfection. | The team of Technion 2022 used this composite as a part of their cell line measurement system called Oracell. This biological system quantifies their manufactured metabolite-decursin by utilizing its effect on the Hippo pathway which is a signaling pathway that regulated proliferation and apoptosis in mammalian cells [1]. TEAD transcriptional factors bind to the 8XGTIIC motifs and regulate the expression of the luciferase gene which is cloned downstream to these motifs. Luciferase is used as a reporter gene that emits luminescence. Thus, the quantity of this metabolite is measured according to the luminescence values, by using a calibration curve. This composite is transfected to CHO (Chinese Hamster Ovarian) cells by stable integration. The role of the Blasticidin S deaminase here is to accomplish a successful selection for positive cells after transfection. | ||
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1.Yu, F. X., & Guan, K. L. (2013). The Hippo pathway: Regulators and regulations. In Geneand Development (Vol. 27, Issue 4, pp. 355–371). doi: 10.1101/gad.210773.112 | 1.Yu, F. X., & Guan, K. L. (2013). The Hippo pathway: Regulators and regulations. In Geneand Development (Vol. 27, Issue 4, pp. 355–371). doi: 10.1101/gad.210773.112 | ||
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| − | <span class='h3bb'>Sequence and Features</span> | + | <span class='h3bb'> |
| + | Sequence and Features</span> | ||
<partinfo>BBa_K4179020 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4179020 SequenceAndFeatures</partinfo> | ||
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<partinfo>BBa_K4179020 parameters</partinfo> | <partinfo>BBa_K4179020 parameters</partinfo> | ||
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Revision as of 05:54, 8 October 2022
Luc_Blast_OraCell
The composite part comprises a Blasticidin S deaminase (BBa_K1943015), which is a Blasticidin resistance gene with the Kozak sequence. Downstream to this under the CMV (BBa_I712004) which is a constitutive promoter, an SV40PolyA terminator (BBa_K4235020) is cloned. Together in this composite, a cTNT promoter (BBa_K4179021) is found, downstream to the 8XGTIIC motifs (BBa_K4179017) followed by the luciferase gene (BBa_I712019) and the low most part SV40 Poly(A)(BBa_K4235020).
Usage and Biology
The team of Technion 2022 used this composite as a part of their cell line measurement system called Oracell. This biological system quantifies their manufactured metabolite-decursin by utilizing its effect on the Hippo pathway which is a signaling pathway that regulated proliferation and apoptosis in mammalian cells [1]. TEAD transcriptional factors bind to the 8XGTIIC motifs and regulate the expression of the luciferase gene which is cloned downstream to these motifs. Luciferase is used as a reporter gene that emits luminescence. Thus, the quantity of this metabolite is measured according to the luminescence values, by using a calibration curve. This composite is transfected to CHO (Chinese Hamster Ovarian) cells by stable integration. The role of the Blasticidin S deaminase here is to accomplish a successful selection for positive cells after transfection.
References
1.Yu, F. X., & Guan, K. L. (2013). The Hippo pathway: Regulators and regulations. In Geneand Development (Vol. 27, Issue 4, pp. 355–371). doi: 10.1101/gad.210773.112 Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 213
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1090