Difference between revisions of "Part:BBa K5206003"

 
 
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<partinfo>BBa_K5206003 SequenceAndFeatures</partinfo>
 
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==Results==
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===Plasmid construction===
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The plasmid vector pETDuet-1 is used to construct a recombinant plasmid pETDuet-1-PalsI-alsR to verify the effect of BBa_K5206003. We amplified alsR and the PalsI region using primers alsR-For and alsR-Rev, and amplified the vector using pETDuet1-gj(alsR)-For and pETDuet1-gj(alsR)-Rev. Subsequently, Gibson assembly was used to ligate the vector and the target gene fragment.
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  <div class="unterschrift"><b> pETDuet-1-PalsI-alsR </b>
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  <div class="unterschrift"><b> Electrophoretic detection of PCR </b>
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K5206003 parameters</partinfo>
 
<partinfo>BBa_K5206003 parameters</partinfo>
 
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Latest revision as of 19:32, 30 September 2024


Palsl-alsR

This part is composed of BBa_K5206000, BBa_K5206001 and BBa_K731721. BBa_K5206000 (alsR) is derived from Escherichia coli BL21(DE3), which is terminated by BBa_K731721 (T7 terminator). In the E. coli genome, AlsR is responsible for the control of six genes related to D-allose metabolism, aISRBACEK and its bidirectional promoter BBa_K5206001(PalsI). The gene downstream of the bidirectional promoter is also regulated by AlsR. When D-allose is present in the environment, the AslR transcriptional regulatory protein binds preferentially to D-allose and deregulates downstream genes.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1263
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Results

Plasmid construction

The plasmid vector pETDuet-1 is used to construct a recombinant plasmid pETDuet-1-PalsI-alsR to verify the effect of BBa_K5206003. We amplified alsR and the PalsI region using primers alsR-For and alsR-Rev, and amplified the vector using pETDuet1-gj(alsR)-For and pETDuet1-gj(alsR)-Rev. Subsequently, Gibson assembly was used to ligate the vector and the target gene fragment.

pETDuet-1-PalsI-alsR

Electrophoretic detection of PCR