Difference between revisions of "Part:BBa K567008"

 
 
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__NOTOC__
 
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<partinfo>BBa_K567008 short</partinfo>
 
<partinfo>BBa_K567008 short</partinfo>
  
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This biobrick is constructed by putting modified enzyme luciferase under promoter T7 and controlled by lac operator. 2 AGG codons and 2 GCG codons are inserted after the ATG start codon of wild type luciferase (BBa_I712019). Modified luciferase keeps the activity of converting luciferin into oxyluciferin, during which bioluminescence will emit. This part is one of the reporter genes to testify the influence of different number of rare codons in regulating protein biosynthesis. This part is used as a measurement to testify the function of LacI -Ptac-tRNA(Arg) (BBa_K567001) or sulA promoter-tRNA(Arg) (BBa_K567002).  
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This biobrick is constructed by putting modified enzyme luciferase under promoter T7 and controlled by lac operator. 2 AGG codons and 2 GCG codons are inserted after the ATG start codon of wild type luciferase (BBa_I712019). Modified luciferase keeps the activity of converting luciferin into oxyluciferin, during which bioluminescence will emit. This part is one of the reporter genes to testify the influence of different number of rare codons in regulating protein biosynthesis. This part is used as a measurement to testify the function of LacI -Ptrc-tRNA(Arg) (BBa_K567001) or sulA promoter-tRNA(Arg) (BBa_K567002).  
 
Cell is cultured in 50ug/ml kanamycin 10ug/ml tetracycline LB liquid medium. When the OD600 of the culture reaches 0.3 IPTG is added to make the final concentration 0.5nM to induce the synthesis of tRNA.  
 
Cell is cultured in 50ug/ml kanamycin 10ug/ml tetracycline LB liquid medium. When the OD600 of the culture reaches 0.3 IPTG is added to make the final concentration 0.5nM to induce the synthesis of tRNA.  
 
Ultrasonication is used to release the luciferase from the cell. Sonics ON 3 seconds, OFF 3 seconds, total ultrasonication time 3minutes.  
 
Ultrasonication is used to release the luciferase from the cell. Sonics ON 3 seconds, OFF 3 seconds, total ultrasonication time 3minutes.  
 
Amount of bioluminescence produced can be detected using luminometer.
 
Amount of bioluminescence produced can be detected using luminometer.
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'''Get more information from Biobrick P''bla''-Luc-2AGG    ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567004 BBa_K567004])'''
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=='''Related Biobrick'''==
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''lacI''-Ptrc-tRNA<sup>Arg</sup>  ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567001 BBa_K567001])
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P''bla''-Luc-2AGG    ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567004 BBa_K567004])
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P''bla''-Luc-4AGG    ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567005 BBa_K567005])
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P''bla''-Luc-6AGG    ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567006 BBa_K567006])
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P''bla''-Luc-8AGG    ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567007 BBa_K567007])
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PT7-Luc-2AGG        ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567008 BBa_K567008])
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PT7-Luc-4AGG        ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567009 BBa_K567009])
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PT7-Luc-6AGG        ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567019 BBa_K567019])
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PT7-Luc-8AGG        ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567010 BBa_K567010])
  
  
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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==Functional Parameters: Austin_UTexas==
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<html>
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<body>
 
<partinfo>BBa_K567008 parameters</partinfo>
 
<partinfo>BBa_K567008 parameters</partinfo>
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<!-- -->
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<h3><center>Burden Imposed by this Part:</center></h3>
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<figure>
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<div class = "center">
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<center><img src = "https://static.igem.org/mediawiki/parts/f/fa/T--Austin_Utexas--no_burden_icon.png" style = "width:160px;height:120px"></center>
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</div>
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<figcaption><center><b>Burden Value: -1.2 ± 2.1% </b></center></figcaption>
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</figure>
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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 BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. 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, an explanation of the sources of burden,  and other conclusions from a large-scale measurement project conducted by the <a href="http://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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</body>
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</html>

Latest revision as of 04:25, 25 August 2020

PT7-Luc-2AGG

This biobrick is constructed by putting modified enzyme luciferase under promoter T7 and controlled by lac operator. 2 AGG codons and 2 GCG codons are inserted after the ATG start codon of wild type luciferase (BBa_I712019). Modified luciferase keeps the activity of converting luciferin into oxyluciferin, during which bioluminescence will emit. This part is one of the reporter genes to testify the influence of different number of rare codons in regulating protein biosynthesis. This part is used as a measurement to testify the function of LacI -Ptrc-tRNA(Arg) (BBa_K567001) or sulA promoter-tRNA(Arg) (BBa_K567002). Cell is cultured in 50ug/ml kanamycin 10ug/ml tetracycline LB liquid medium. When the OD600 of the culture reaches 0.3 IPTG is added to make the final concentration 0.5nM to induce the synthesis of tRNA. Ultrasonication is used to release the luciferase from the cell. Sonics ON 3 seconds, OFF 3 seconds, total ultrasonication time 3minutes. Amount of bioluminescence produced can be detected using luminometer.

Get more information from Biobrick Pbla-Luc-2AGG (BBa_K567004)

Related Biobrick

lacI-Ptrc-tRNAArg (BBa_K567001)

Pbla-Luc-2AGG (BBa_K567004)

Pbla-Luc-4AGG (BBa_K567005)

Pbla-Luc-6AGG (BBa_K567006)

Pbla-Luc-8AGG (BBa_K567007)

PT7-Luc-2AGG (BBa_K567008)

PT7-Luc-4AGG (BBa_K567009)

PT7-Luc-6AGG (BBa_K567019)

PT7-Luc-8AGG (BBa_K567010)


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 48
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 48
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 48
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 911


Functional Parameters: Austin_UTexas

BBa_K567008 parameters

Burden Imposed by this Part:

Burden Value: -1.2 ± 2.1%

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 BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods, an explanation of the sources of burden, 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.