Difference between revisions of "Part:BBa K2888006"

 
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<partinfo>BBa_K2888006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2888006 SequenceAndFeatures</partinfo>
  
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===Introduction===
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We have designed, based on our original goal, a basic part of chimeric lysozyme and mlrA gene with a linker sequence in between in order to create an enzyme that retain the both the function of lysing the cyanobacteria and denaturing their toxin MCLR. This part is intended to resolve the ultimate problem of cyanobacteria pollution. Therefore, the composite part contains our promoter, RBS, 6✖️His tag, lysozyme gene, a linker in between, mlrA gene and a terminator. During some of the experimental trials, we also managed to add a Sumo tag before the  6✖️His tag in order to increase the solubility of the enzyme.
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===Experience===
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Through Nested PCR, we are able to successfully infuse the lysozyme- mlrA gene into the PSB1C3 backbone. The linearized plasmid after the infusion is seen below on lane 2 and 8 of the gel. However, due to limited time, we were only able to focus on the function of lysozyme, which is part BBa_K2888002 and part BBa_K2888003.
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<img src="https://static.igem.org/mediawiki/parts/f/f1/T--SBS_SH_112144--plasmid.png" width=400 height=200/>
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<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 23:25, 17 October 2018


Chimerical gene (lysozyme and mlrA)

This part is the fusion of two basic parts in order to combine two separate function simultaneously: lyse cyanobacterial wall and degrade MCLR simultaneously.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1288
    Illegal AgeI site found at 823
  • 1000
    COMPATIBLE WITH RFC[1000]

Introduction

We have designed, based on our original goal, a basic part of chimeric lysozyme and mlrA gene with a linker sequence in between in order to create an enzyme that retain the both the function of lysing the cyanobacteria and denaturing their toxin MCLR. This part is intended to resolve the ultimate problem of cyanobacteria pollution. Therefore, the composite part contains our promoter, RBS, 6✖️His tag, lysozyme gene, a linker in between, mlrA gene and a terminator. During some of the experimental trials, we also managed to add a Sumo tag before the 6✖️His tag in order to increase the solubility of the enzyme.


Experience

Through Nested PCR, we are able to successfully infuse the lysozyme- mlrA gene into the PSB1C3 backbone. The linearized plasmid after the infusion is seen below on lane 2 and 8 of the gel. However, due to limited time, we were only able to focus on the function of lysozyme, which is part BBa_K2888002 and part BBa_K2888003.