Difference between revisions of "Part:BBa K1689011"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1689011 short</partinfo> | <partinfo>BBa_K1689011 short</partinfo> | ||
− | + | dCas9 fused with Δα segment of β-glactosidase | |
For their ease of use, low assay time, cost-effectiveness, high sensitivity, electrochemical biosensors have emerged as an attractive method for molecular detection. Combing with our PC Reporter we may further enhance it's applicability, that is to implement point-of-care diagnostics in settings outside clinics and without the need of specialized analytical laboratories. | For their ease of use, low assay time, cost-effectiveness, high sensitivity, electrochemical biosensors have emerged as an attractive method for molecular detection. Combing with our PC Reporter we may further enhance it's applicability, that is to implement point-of-care diagnostics in settings outside clinics and without the need of specialized analytical laboratories. | ||
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In our project, we could dissect LacZ as Δα, ∆ω segments to fuse with dCas9 to do the same as our PC Reporter. | In our project, we could dissect LacZ as Δα, ∆ω segments to fuse with dCas9 to do the same as our PC Reporter. | ||
β-galactosidase is used to catalyze the hydrolysis of p-aminophenyl-β-D-galactopyranoside (PAPG) to p-aminophenol (PAP), which can be oxidized at an working electrode held at 220mV versus the reference electrode. | β-galactosidase is used to catalyze the hydrolysis of p-aminophenyl-β-D-galactopyranoside (PAPG) to p-aminophenol (PAP), which can be oxidized at an working electrode held at 220mV versus the reference electrode. | ||
+ | [[File: https://static.igem.org/mediawiki/2015/a/af/Peking-Hardware-From_Bioluminescence_to_Electronic_signal-lactamase.png]] | ||
+ | References: | ||
+ | 1 Mohler, W. A., & Blau, H. M. (1996). Gene expression and cell fusion analyzed by lacZ complementation in mammalian cells. Proceedings of the National Academy of Sciences, 93(22), 12423-12427. | ||
+ | 2 | ||
+ | 3 Biran, I., Klimentiy, L., Hengge-Aronis, R., Ron, E. Z., & Rishpon, J. (1999). On-line monitoring of gene expression. Microbiology, 145(8), 2129-2133. | ||
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Revision as of 16:59, 18 September 2015
dCas9-delta alpha dCas9 fused with Δα segment of β-glactosidase
For their ease of use, low assay time, cost-effectiveness, high sensitivity, electrochemical biosensors have emerged as an attractive method for molecular detection. Combing with our PC Reporter we may further enhance it's applicability, that is to implement point-of-care diagnostics in settings outside clinics and without the need of specialized analytical laboratories.
β-glactosidase (LacZ) is an exoglycosidase from E.coli, composing of 1024 amino acids. LacZ is a commonly used as a reporter, known for blue white screen, which catalyzes X-gal to produce blue dye; by using different substrates, for example ONPG, reaction can be measured quantitatively.
In our project, we could dissect LacZ as Δα, ∆ω segments to fuse with dCas9 to do the same as our PC Reporter. β-galactosidase is used to catalyze the hydrolysis of p-aminophenyl-β-D-galactopyranoside (PAPG) to p-aminophenol (PAP), which can be oxidized at an working electrode held at 220mV versus the reference electrode. File:Https://static.igem.org/mediawiki/2015/a/af/Peking-Hardware-From Bioluminescence to Electronic signal-lactamase.png References: 1 Mohler, W. A., & Blau, H. M. (1996). Gene expression and cell fusion analyzed by lacZ complementation in mammalian cells. Proceedings of the National Academy of Sciences, 93(22), 12423-12427. 2 3 Biran, I., Klimentiy, L., Hengge-Aronis, R., Ron, E. Z., & Rishpon, J. (1999). On-line monitoring of gene expression. Microbiology, 145(8), 2129-2133.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1150
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 4
Illegal BamHI site found at 3429 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]