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> | ||
+ | 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. | ||
+ | <p style="text-align: center!important;">https://static.igem.wiki/teams/4891/wiki/parts/6.png</p> | ||
+ | <p style="text-align: center!important;"><b>Figure 1 Knock out of aroK gene.</b></p> | ||
+ | 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> | ||
+ | <p style="text-align: center!important;"><b>Figure 2 Knock out of aroL gene.</b></p> | ||
+ | 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> | ||
+ | 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> | ||
+ | <p style="text-align: center!important;"><b>Figure 3 Growth curve in the engineered strains without (left) or with amino acid (right).</b></p> | ||
+ | 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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 200