Difference between revisions of "Part:BBa K422014"

Revision as of 10:31, 27 October 2010

mCyPet (AarI A-part)

Biological Background

CyPet is an evolutionary optimized cyan fluorescent protein CFP developed in 2005 by Nguyen and Daugherty. They showed the FRET pair CyPet-YPet to have a 7-fold higher ratiometric FRET signal change than their parental pair.

mCyPet has a mutation in A206K preventing it from forming dimers (Ohashi et al., 20007).

Characterization

Figure 1: Fluorescence spectra of mCyPet measured at an excitation wavelength of 425 nm.

mCyPet has a distinct fluorescence spectra. Measured at an excitation wavelength of 425 nm with a Fluorescein High Precision Monochromator, maximum emission is detected at 482 nm (Figure 1).

Cloning strategy

BBF RFC28: A method for combinatorial multi-part assembly based on the Type IIs restriction enzyme AarI. mcyPet-A is an A-part compatible for N-Terminal fusion. For more information see [http://2010.igem.org/Team:ETHZ_Basel/Biology/Cloning].

Sequence and Features