Difference between revisions of "Part:BBa K3875006"
 
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<i>trpG</i> (<a href = "https://parts.igem.org/Part:BBa_K3875003" > BBa_K3875003</a>),  
 
<i>trpG</i> (<a href = "https://parts.igem.org/Part:BBa_K3875003" > BBa_K3875003</a>),  
 
<i>trpC</i> (<a href = "https://parts.igem.org/Part:BBa_K3875002" > BBa_K3875002</a>).<br>
 
<i>trpC</i> (<a href = "https://parts.igem.org/Part:BBa_K3875002" > BBa_K3875002</a>).<br>
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This part could express two enzymes, indole-3-glycerol-phosphate synthase (IGPS, encoded by <i>trpC</i>) and anthranilate synthase (AS, encoded by <i>trpE</i>). <br>
 
This part could express two enzymes, indole-3-glycerol-phosphate synthase (IGPS, encoded by <i>trpC</i>) and anthranilate synthase (AS, encoded by <i>trpE</i>). <br>
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Erythritos-4-phosphate (E4P) from the pentose phosphate pathway (<strong>PPP</strong>) is condensed with <strong>phosphoenolpyruvate (PEP)</strong> from the glycolysis pathway <strong>(EMP)</strong> to form 3-deoxy-D-arabinheptanose-7-phosphate <strong>(DAHP)</strong>. <br>
 
Erythritos-4-phosphate (E4P) from the pentose phosphate pathway (<strong>PPP</strong>) is condensed with <strong>phosphoenolpyruvate (PEP)</strong> from the glycolysis pathway <strong>(EMP)</strong> to form 3-deoxy-D-arabinheptanose-7-phosphate <strong>(DAHP)</strong>. <br>
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After 7 steps, DAHP is converted to chorismate. Our <i>trpE</i> and <i>trpC</i> genes in <i>E. coli</i> promote the chorismate into the latter half of the 5-HTP synthesis pathway <strong>(Figure 1)</strong>.<br>
 
After 7 steps, DAHP is converted to chorismate. Our <i>trpE</i> and <i>trpC</i> genes in <i>E. coli</i> promote the chorismate into the latter half of the 5-HTP synthesis pathway <strong>(Figure 1)</strong>.<br>
  
 
<img src="https://static.igem.org/mediawiki/parts/7/72/T--BUCT--_Reaction_of_trpE_and_trpC.png"width="640px";height="30px"/>
 
<img src="https://static.igem.org/mediawiki/parts/7/72/T--BUCT--_Reaction_of_trpE_and_trpC.png"width="640px";height="30px"/>
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<img src="https://static.igem.org/mediawiki/parts/c/c4/T--BUCT--_Plasmid_map.png"width="640px";height="30px"/><br>
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<strong>Below is plasmid profile of pSA74-lac-trpE-trpG-AntrpC-T1</strong>
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<img src="https://static.igem.org/mediawiki/parts/3/3b/T--BUCT--_Plasmid_map.jpg"width="640px";height="30px"/><br>
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<strong>Method</strong>:
 
<strong>Method</strong>:
 
The <i>pcd</i> and <i>p4h</i> genes were amplified by <strong>PCR</strong>, then the two genes and pSA74 were digested by restriction endonuclease, and finally <strong>pSA74-lac-trpE-trpG-AntrpC-T1</strong> was obtained by ligase T4. Then HindIII and BamHI were used for digestion verification.<br>
 
The <i>pcd</i> and <i>p4h</i> genes were amplified by <strong>PCR</strong>, then the two genes and pSA74 were digested by restriction endonuclease, and finally <strong>pSA74-lac-trpE-trpG-AntrpC-T1</strong> was obtained by ligase T4. Then HindIII and BamHI were used for digestion verification.<br>

Latest revision as of 09:14, 18 October 2021


Usage and Biology

Design: Contained gene: trpE ( BBa_K3875009), trpG ( BBa_K3875003), trpC ( BBa_K3875002).
This part could express two enzymes, indole-3-glycerol-phosphate synthase (IGPS, encoded by trpC) and anthranilate synthase (AS, encoded by trpE).
Erythritos-4-phosphate (E4P) from the pentose phosphate pathway (PPP) is condensed with phosphoenolpyruvate (PEP) from the glycolysis pathway (EMP) to form 3-deoxy-D-arabinheptanose-7-phosphate (DAHP).
After 7 steps, DAHP is converted to chorismate. Our trpE and trpC genes in E. coli promote the chorismate into the latter half of the 5-HTP synthesis pathway (Figure 1).






Below is plasmid profile of pSA74-lac-trpE-trpG-AntrpC-T1




Method: The pcd and p4h genes were amplified by PCR, then the two genes and pSA74 were digested by restriction endonuclease, and finally pSA74-lac-trpE-trpG-AntrpC-T1 was obtained by ligase T4. Then HindIII and BamHI were used for digestion verification.
Result: The result of agarose gel electrophoresis proves that our plasmid has been successfully constructed .

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 600
    Illegal BglII site found at 661
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 4128
  • 1000
    COMPATIBLE WITH RFC[1000]