Difference between revisions of "Part:BBa K4891006"
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===Usage and Biology===
 
===Usage and Biology===
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<h3>1 PCR verification</h3>
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Colony PCR results show that aroK and aroL genes have been knocked out from the genome of the host strain (Figures 1-2), and the recombinant strains are named as strains E. coli YCY2-E. coli YCY3, respectively.
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<p style="text-align: center!important;">https://static.igem.wiki/teams/4891/wiki/parts/6.png</p>
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<p style="text-align: center!important;"><b>Figure 1 Knock out of aroK gene.</b></p>
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The DNA fragment of knocked out and un-knocked out for aroK gene are 1800 bp and 2322 bp, respectively.
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<p style="text-align: center!important;">https://static.igem.wiki/teams/4891/wiki/parts/7.png</p>
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<p style="text-align: center!important;"><b>Figure 2 Knock out of aroL gene.</b></p>
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The DNA fragment of knocked out and un-knocked out for aroL gene are 1800 bp and 2325 bp, respectively.
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<h3>2 Growth assay</h3>
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To evaluate whether the elimination of shikimate catabolism resulted in an auxotroph for amino acids, the growth curve in strains E. coli YCY1-YCY3 is examined. As seen in Figure 3, the growth rate and glucose consumption of strain E. coli YCY2-3 are significantly slowed down without the addition of L-tyrosine, L-phenylalanine, and L-tryptophan. In contrast, the growth status of these strains is improved after supplementing aromatic amino acids.
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<p style="text-align: center!important;">https://static.igem.wiki/teams/4891/wiki/parts/8.png</p>
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<p style="text-align: center!important;"><b>Figure 3 Growth curve in the engineered strains without (left) or with amino acid (right).</b></p>
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WT (Escherichia coli MG1655), YCY1 (MG1655 ΔldhA ΔadhE ΔpoxB Δpta), YCY2 (MG1655 ΔldhA ΔadhE ΔpoxB Δpta ΔaroK), YCY3 (MG1655 ΔldhA ΔadhE ΔpoxB Δpta ΔaroK ΔaroL)
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Revision as of 08:05, 8 October 2023


aroK

With the disruption of shikimate catabolism, shikimate production could be improved.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 200