Difference between revisions of "Part:BBa K4891014"
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<partinfo>BBa_K4891014 short</partinfo> | <partinfo>BBa_K4891014 short</partinfo> | ||
− | It is the key part that is responsible for strengthening glucose utilization rate. The phosphotransferase system was replaced by heterogeneously introducing glucose facilitator in strain E. coli ptsG | + | It is the key part that is responsible for strengthening glucose utilization rate. The phosphotransferase system was replaced by heterogeneously introducing glucose facilitator in strain E. coli ptsG:glk-glf. |
− | <!-- Add more about the biology of this part here | + | <!-- Add more about the biology of this part here--> |
===Usage and Biology=== | ===Usage and Biology=== | ||
+ | We engineered the shikimate pathway of E. coli MG1655 to efficiently accumulate SA. We knocked out ptsG, ldhA, adhE, poxB, and pta genes to achieve the production of precursor substance PEP. To increase intracellular E4P content, we overexpressed tktA and talB genes. To enhance product accumulation, we overexpressed aroG, aroB, aroD, and aroE genes, while knocking down aroK and aroL genes to cut off the metabolic flux, thus accomplishing the accumulation of shikimic acid. Besides, a non-phosphorylated pathway, that is, glk and glf genes (mainly by glk-glf integration into the ptsG locus) is introduced to enhance glucose utilization. To achieve the goal above-mentioned, we totally constructed 26 parts this year. See table below: | ||
+ | {| | ||
+ | | <html><img style="width:400px" src="https://static.igem.wiki/teams/4891/wiki/parts/22.png" ></html> | ||
+ | |- | ||
+ | |} | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> |
Latest revision as of 15:10, 9 October 2023
glk-glf
It is the key part that is responsible for strengthening glucose utilization rate. The phosphotransferase system was replaced by heterogeneously introducing glucose facilitator in strain E. coli ptsG:glk-glf.
Usage and Biology
We engineered the shikimate pathway of E. coli MG1655 to efficiently accumulate SA. We knocked out ptsG, ldhA, adhE, poxB, and pta genes to achieve the production of precursor substance PEP. To increase intracellular E4P content, we overexpressed tktA and talB genes. To enhance product accumulation, we overexpressed aroG, aroB, aroD, and aroE genes, while knocking down aroK and aroL genes to cut off the metabolic flux, thus accomplishing the accumulation of shikimic acid. Besides, a non-phosphorylated pathway, that is, glk and glf genes (mainly by glk-glf integration into the ptsG locus) is introduced to enhance glucose utilization. To achieve the goal above-mentioned, we totally constructed 26 parts this year. See table below:
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Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 124
Illegal PstI site found at 1136
Illegal PstI site found at 1327
Illegal PstI site found at 1900 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 124
Illegal PstI site found at 1136
Illegal PstI site found at 1327
Illegal PstI site found at 1900 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 124
Illegal PstI site found at 1136
Illegal PstI site found at 1327
Illegal PstI site found at 1900 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 124
Illegal PstI site found at 1136
Illegal PstI site found at 1327
Illegal PstI site found at 1900
Illegal AgeI site found at 421
Illegal AgeI site found at 1312
Illegal AgeI site found at 1468
Illegal AgeI site found at 1480
Illegal AgeI site found at 2191 - 1000COMPATIBLE WITH RFC[1000]