Difference between revisions of "Part:BBa K4368004"
 
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<partinfo>BBa_K4368004 short</partinfo>
 
<partinfo>BBa_K4368004 short</partinfo>
  
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''GlgC16'' encodes for the ADP-glucose pyrophosphorylase gene of ''Escherichia coli'' ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K118016 BBa_K118016]). This enzyme is responsible for the synthesis of starch-like starch using glucose as a substrate. In addition, this part includes the composition used by the team, which includes a strong rbs ([https://parts.igem.org/wiki/index.php?title=Part:BBa_B0030 BBa_B0030]), a double terminator ([https://parts.igem.org/wiki/index.php?title=Part:BBa_B0015 BBa_B0015]) as well as a promoter inducible by the cI protein of the phage lambda ([https://parts.igem.org/wiki/index.php?title=Part:BBa_R1051 BBa_R1051]).
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==Description==
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The gene has been placed under the control of this promoter to build the glucose concentration-based gene regulatory circuit that integrates all our parts.
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''GlgC16'' encodes for the ADP-glucose pyrophosphorylase gene of ''Escherichia coli'' (<partinfo>BBa_K118016</partinfo>). This enzyme is responsible for the synthesis of starch-like starch using glucose as a substrate. In addition, this part includes the composition used by the team, which includes a strong rbs (<partinfo>BBa_B0030</partinfo>), a double terminator (<partinfo>BBa_B0015</partinfo>) as well as a promoter inducible by the ''cI'' protein of the phage lambda (<partinfo>BBa_R1051</partinfo>).
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<partinfo>BBa_K118016</partinfo> works under the control of this promoter to build the glucose concentration-based gene regulatory circuit that integrates all our parts.
  
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==Characterization==
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The expression cassette sequence was digested with EcoRI and PstI enzymes and subsequently ligated with a chloramphenicol resistant plasmid backbone (Cm).
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Transforming bacteria were created with this plasmid and seeded on LB-Agar+Cm plates. After growth, colonies were selected based on their color (white) and DNA extraction was performed using the Promega PureYield Plasmid Miniprep System kit.
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The resulting DNA is used for further digestion with EcoRI and PstI. The digests are then run on a 0.75% agarose gel at 90 mV voltage and constant amperage. BioRad brand RedSafe is used as an intercalating developing agent.
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===Enzyme digestion===
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[[File:glgC.png|350px|center|]]
  
 
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<span class='h3bb'>Sequence and Features</span>
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==<span class='h3bb'>Sequence and Features</span>==
 
<partinfo>BBa_K4368004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4368004 SequenceAndFeatures</partinfo>
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===Characterization from iGEM22_UMA_MALAGA===
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==Contribution ==
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*'''Group:''' [https://2022.igem.wiki/uma-malaga/index.html UMA_MALAGA]
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*'''Author:''' Jiménez Amores, Carmen
  
 
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Latest revision as of 18:54, 7 October 2022


pcI + rbs + glgC16 + terminator

Description

GlgC16 encodes for the ADP-glucose pyrophosphorylase gene of Escherichia coli (BBa_K118016). This enzyme is responsible for the synthesis of starch-like starch using glucose as a substrate. In addition, this part includes the composition used by the team, which includes a strong rbs (BBa_B0030), a double terminator (BBa_B0015) as well as a promoter inducible by the cI protein of the phage lambda (BBa_R1051). BBa_K118016 works under the control of this promoter to build the glucose concentration-based gene regulatory circuit that integrates all our parts.

Characterization

The expression cassette sequence was digested with EcoRI and PstI enzymes and subsequently ligated with a chloramphenicol resistant plasmid backbone (Cm). Transforming bacteria were created with this plasmid and seeded on LB-Agar+Cm plates. After growth, colonies were selected based on their color (white) and DNA extraction was performed using the Promega PureYield Plasmid Miniprep System kit. The resulting DNA is used for further digestion with EcoRI and PstI. The digests are then run on a 0.75% agarose gel at 90 mV voltage and constant amperage. BioRad brand RedSafe is used as an intercalating developing agent.

Enzyme digestion

GlgC.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 272
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Contribution