Difference between revisions of "Part:BBa K422014"

Revision as of 10:33, 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.

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 shows the relative fluorescene units of mCyPet over a wavelenght range of 460 to 560 nm.

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

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal SpeI site found at 733
12
INCOMPATIBLE WITH RFC[12]
Illegal SpeI site found at 733
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal SpeI site found at 733
25
INCOMPATIBLE WITH RFC[25]
Illegal SpeI site found at 733
Illegal NgoMIV site found at 697
1000
COMPATIBLE WITH RFC[1000]

@@ Line 9: / Line 9: @@
 ===Characterization===
-[[Image:mCyPet.png|thumb|200px|'''Figure 1:''' Fluorescence spectra of mCyPet measured at an excitation wavelength of 425 nm.]]
+[[Image:mCyPet.png|thumb|200px|'''Figure 1:''' Fluorescence spectra of mCyPet.]]
-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).
+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 shows the relative fluorescene units of mCyPet over a wavelenght range of 460 to 560 nm.
 ===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].
+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].