Difference between revisions of "Part:BBa K660004"

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Figure 1: Digram showing movement of TMV (tobacco mosaic virus). On the left shows TMV with iLOV, and the centre and right show TMV with GFP. We can see that TMViLOV shows systemic infection, whereas TMVGFP shows poor, or no infection.Image taken from: Chapman, S. et al (2008) The photo-reversible fluorescent protein iLOV outperforms GFP as a reporter of plant virus infection. PNAS, 105 (50) pp. 20038 - 20043
 
Figure 1: Digram showing movement of TMV (tobacco mosaic virus). On the left shows TMV with iLOV, and the centre and right show TMV with GFP. We can see that TMViLOV shows systemic infection, whereas TMVGFP shows poor, or no infection.Image taken from: Chapman, S. et al (2008) The photo-reversible fluorescent protein iLOV outperforms GFP as a reporter of plant virus infection. PNAS, 105 (50) pp. 20038 - 20043
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(Full article available at: http://www.pnas.org/content/105/50/20038.full)
  
  
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Figure 2: Image demonstrating photo-bleaching reversibility of iLOV. Left – iLOV pre-bleach, Centre – iLOV post-bleach, and Right – iLOV post-recovery. Image taken from Chapman et al (2008) The photo-reversible fluorescent protein iLOV outperforms GFP as a reporter of plant virus infection. PNAS, 105 (50) pp. 20038 - 20043
 
Figure 2: Image demonstrating photo-bleaching reversibility of iLOV. Left – iLOV pre-bleach, Centre – iLOV post-bleach, and Right – iLOV post-recovery. Image taken from Chapman et al (2008) The photo-reversible fluorescent protein iLOV outperforms GFP as a reporter of plant virus infection. PNAS, 105 (50) pp. 20038 - 20043
 +
(Full article available at: http://www.pnas.org/content/105/50/20038.full)
  
 
Another advantage of LOV in comparison to other fluorescent proteins is it's ability to work in anoxic conditions, making it ideally suited to work within biofilms.
 
Another advantage of LOV in comparison to other fluorescent proteins is it's ability to work in anoxic conditions, making it ideally suited to work within biofilms.

Revision as of 13:23, 21 September 2011

iLOV

iLOV is a a variant of LOV2 (https://parts.igem.org/Part:BBa_K660000) which has been isolated over time as it shows better fluorescence and photo-stabiltility.

In studies whereby it has been used in comparison to GFP to track plant infections, it has outperformed GFP. It is much smaller than GFP at around 300 bp as opposed to 700 bp, and for this reason it better suited as a reporter for the movement of things such as viruses, and tagging of small proteins. Unlike GFP which photo-bleaches irreversibly, iLOV undergoes spontaneous recovery from photo-bleaching under high intensity exposure to UV. This photo-bleaching reversibility is due to the changing state between the fluorescent and non-fluorescent form of the bound FMN chromopore

"Ilovversusgfp.jpg

Figure 1: Digram showing movement of TMV (tobacco mosaic virus). On the left shows TMV with iLOV, and the centre and right show TMV with GFP. We can see that TMViLOV shows systemic infection, whereas TMVGFP shows poor, or no infection.Image taken from: Chapman, S. et al (2008) The photo-reversible fluorescent protein iLOV outperforms GFP as a reporter of plant virus infection. PNAS, 105 (50) pp. 20038 - 20043 (Full article available at: http://www.pnas.org/content/105/50/20038.full)


Ilovephotobleaching.JPG

Figure 2: Image demonstrating photo-bleaching reversibility of iLOV. Left – iLOV pre-bleach, Centre – iLOV post-bleach, and Right – iLOV post-recovery. Image taken from Chapman et al (2008) The photo-reversible fluorescent protein iLOV outperforms GFP as a reporter of plant virus infection. PNAS, 105 (50) pp. 20038 - 20043 (Full article available at: http://www.pnas.org/content/105/50/20038.full)

Another advantage of LOV in comparison to other fluorescent proteins is it's ability to work in anoxic conditions, making it ideally suited to work within biofilms. Fluorescent imaging of iLOV can be done by using an excitation wavelength of 476nm, with fluorescent emission between 510 and 550 nm. (Chapman et al 2008)

This protein has been codon optimised for E.coli expression.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]