Part:BBa_K4761223
adh6-linker4-ARO10
adh6-linker4-ARO10 fusion expression unit for tyrosol production. Designed based on BBa_K4761001 and BBa_K4761000 linked with BBa_K4761033, a rigid linker.Because the reaction catalyzed by yeast-derived pyruvate decarboxylase ARO10 and endogenous alcohol dehydrogenase ADH6 is an adjacent reaction in salidroside metabolism, the adh6 gene and ARO10 gene are linked through the linker designed by 2023 HiZJU-China in attempt to increase its generating efficiency.
For guidelines for fusion PCR and guidelines for its primer design, see BBa_K4761222.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2886
Illegal SpeI site found at 1837
Illegal PstI site found at 1715 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2886
Illegal SpeI site found at 1837
Illegal PstI site found at 1715 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2886
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 2886
Illegal SpeI site found at 1837
Illegal PstI site found at 1715 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 2886
Illegal SpeI site found at 1837
Illegal PstI site found at 1715
Illegal NgoMIV site found at 991 - 1000COMPATIBLE WITH RFC[1000]
Characterization
The link was made using a fusion PCR method, in which the stop codon of the adh6 gene was replaced with a linker sequence. For details, see BBa_K4761222.
The fermentation results after fusion expression are shown in Table1 below.
The results show that the rigid joint Linker4 hinders the catalytic efficiency of the reaction, indicating that the rigid joint is not suitable for the improvement of the reaction metabolism.
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