Difference between revisions of "Part:BBa K325218"
 
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<partinfo>BBa_K325218 short</partinfo>
 
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'''Description'''<br>
 
'''Description'''<br>
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This part generates a red-mutant of the luciferase from the Japanese firefly (L.cruciata) as well as the luciferin regenerating enzyme (LRE). It is under the control of an Arabinose induced promoter.
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D-Luciferin has to be added to obtain light output.
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{{:Templates/Cambridge-fireflycolour|colour=orange|substitution=Ser286Asn|id=249}}
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THIS PAGE IS CURRENTLY BEING UPDATED.  
 
THIS PAGE IS CURRENTLY BEING UPDATED.  
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[http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html '''[2&#x5d;:'''] T. Nakatsu ''et al.'' (2006) Structural Basis for the spectral difference in luciferase bioluminescence, ''Nature'' '''440'''(16), 372-376.
 
[http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html '''[2&#x5d;:'''] T. Nakatsu ''et al.'' (2006) Structural Basis for the spectral difference in luciferase bioluminescence, ''Nature'' '''440'''(16), 372-376.
 
[http://www.ncbi.nlm.nih.gov/pubmed/11457857 '''[3&#x5d;:'''] K. Gomi and N. Kajiyama, (2001) Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin, ''The Journal of Biological Chemistry'', '''276'''(39), 36508-36513.
 
[http://www.ncbi.nlm.nih.gov/pubmed/11457857 '''[3&#x5d;:'''] K. Gomi and N. Kajiyama, (2001) Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin, ''The Journal of Biological Chemistry'', '''276'''(39), 36508-36513.
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===Functional Parameters===
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<partinfo>BBa_K325218 parameters</partinfo>
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==Functional Parameters: Austin_UTexas==
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<h3><center>Burden Imposed by this Part:</center></h3>
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<figure>
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<center><img src = "https://static.igem.org/mediawiki/parts/4/43/T--Austin_Utexas--Low_significant_burden.png" style = "width:200px;height:120px"></center>
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<figcaption><center><b>Burden Value: 10.8 ± 7.3% </b></center></figcaption>
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<p> Burden is the percent reduction in the growth rate of <i>E. coli</i> cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This part exhibited a significant burden. Users should be aware that BioBricks with a burden of >20-30% may be susceptible to mutating to become less functional or nonfunctional as an evolutionary consequence of this fitness cost. This risk increases as they used for more bacterial cell divisions or in larger cultures. Users should be especially careful when combining multiple burdensome parts, as plasmids with a total burden of >40% are expected to mutate so quickly that they become unclonable. Refer to any one of the
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<a href="https://parts.igem.org/Part:BBa_K3174002">BBa_K3174002</a> - <a href="https://parts.igem.org/Part:BBa_K3174007">BBa_K3174007</a> pages for more information on the methods and other conclusions from a large-scale measurement project conducted by the <a href="https://2019.igem.org/Team:Austin_UTexas">2019 Austin_UTexas team.</a></p>
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<p>This functional parameter was added by the <a href="https://2020.igem.org/Team:Austin_UTexas/Contribution">2020 Austin_UTexas team.</a></p>
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Latest revision as of 00:31, 4 September 2020

Orange Firefly Luciferase (under pBAD)
L. Cruciata
(E. coli optimised)

Input: L-Arabinose
Output: Light

pBad/araC
I0500		Luciferase/LRE
K325210
 Cambridge-Eglowli.png

Part Main Page        Arabinose -> Light        Add Data       


Description

Input: L-Arabinose
Output: Light (orange)

pBad/araC
I0500		mutated version of
Luciferase/LRE
K325210

This part should show similarity to BBa_K325219     Add Data       


Description
This part is based on part BBa_K325219. Two site-directed mutagenesis events have been carried out on the luciferase, an Asn286 Ser mutation was used to create part BBa_K325209 which is the green wild-type luciferase. Then a Ser286Asn mutation was used to create a orange bioluminescence.


Colour of emission

Figure 1 - Comparison of part BBa_K325249 against other constructs created by the E.glowli team. Note that this picture should be used as a relative comparison rather than absolute colour. All images change markedly depending on the white balance used.


Compatibility
Chassis: Device has been shown to work in Top 10 (Invitrogen)
Plasmids: Device has been shown to work on pSB1C3


THIS PAGE IS CURRENTLY BEING UPDATED.

Pictures

Figure 1 - E.Coli (Invitrogen TOP 10) cells transformed with BBa K325909 (blue light bulb) and BBa 325219 (red light bulb)

</center> 1Measured by the [http://2010.igem.org/Team:Cambridge Cambridge iGEM team 2010]

Compatibility
Chassis: Device has been shown to work in Top 10 (Invitrogen)
Plasmids: Device has been shown to work on pSB1C3


References
[http://www.ncbi.nlm.nih.gov/pubmed/18949818 [1]:] S.M. Marques and J.C.G. Esteves da Silva, (2009) Firefly Bioluminescence: A Mechanistic Approach of Luciferase Catalyzed Reactions,Life 61, 6-17.

[http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html [2]:] T. Nakatsu et al. (2006) Structural Basis for the spectral difference in luciferase bioluminescence, Nature 440(16), 372-376. [http://www.ncbi.nlm.nih.gov/pubmed/11457857 [3]:] K. Gomi and N. Kajiyama, (2001) Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin, The Journal of Biological Chemistry, 276(39), 36508-36513.


Functional Parameters

outputLight
positive_regulatorsL-arabinose



Functional Parameters: Austin_UTexas

Burden Imposed by this Part:

Burden Value: 10.8 ± 7.3%

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This part exhibited a significant burden. Users should be aware that BioBricks with a burden of >20-30% may be susceptible to mutating to become less functional or nonfunctional as an evolutionary consequence of this fitness cost. This risk increases as they used for more bacterial cell divisions or in larger cultures. Users should be especially careful when combining multiple burdensome parts, as plasmids with a total burden of >40% are expected to mutate so quickly that they become unclonable. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.