Difference between revisions of "Part:BBa K1689011"
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In our project, we dissected LacZ as Δα, ∆ω segments to fuse with dCas9 to do the same as our PC Reporter.<br/><br/> | In our project, we dissected LacZ as Δα, ∆ω segments to fuse with dCas9 to do the same as our PC Reporter.<br/><br/> | ||
| − | + | [[File:Peking-speculation-LacZ-1.png|middle| 450px |]] | |
<br/><br/> | <br/><br/> | ||
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.<br/> | 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.<br/> | ||
| − | + | [[File:Peking-speculation-LacZ-2.png|380px|]] | |
| − | + | <br/> | |
References:<br/> | References:<br/> | ||
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.<br/> | 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.<br/> | ||
Revision as of 18:27, 4 November 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. Combined with our PC Reporter, we further enhanced it's applicability, that is to implement pathogen diagnostics in settings outside clinics and without the need of specialized analytical laboratories.
β-glactosidase (LacZ) is an exoglycosidase from E.coli, composed of 1024 amino acids. LacZ is commonly used as 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 dissected 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.
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 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]