Difference between revisions of "Part:BBa K1689011"

Revision as of 17:17, 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.

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