Part:BBa_K3875006
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 600
Illegal BglII site found at 661 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 4128
- 1000COMPATIBLE WITH RFC[1000]
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