Difference between revisions of "Part:BBa K2570001:Design"
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We ordered the part DNA from a synthesis company and had codon optimization of the sequence. | We ordered the part DNA from a synthesis company and had codon optimization of the sequence. | ||
Mutation of restriction sites EcoRⅠ, SpeⅠand PstⅠ. | Mutation of restriction sites EcoRⅠ, SpeⅠand PstⅠ. | ||
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| + | Site-directed mutation site | ||
Mutation 1: Aro10-Pst1 （Pro 619-621）cctgca-ccggca | Mutation 1: Aro10-Pst1 （Pro 619-621）cctgca-ccggca | ||
Revision as of 09:27, 5 October 2018
Aro10
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We ordered the part DNA from a synthesis company and had codon optimization of the sequence. Mutation of restriction sites EcoRⅠ, SpeⅠand PstⅠ.
Site-directed mutation site
Mutation 1: Aro10-Pst1 （Pro 619-621）cctgca-ccggca
Sense primer: TATATTTTTGTTCCGGCAGATTTTG
Anti-sense primer: AATCTGCCGGAACAAAAATATAACC
Mutation 2: Aro10-Spe1 （Thr 741-744）actagt- accagt
Sense primer: ACAAGATTACCAGTTGGATATATTC
Anti-sense primer: ATATATCCAACTGGTAATCTTGTTG
Mutation 3: Aro10-EcoR1（Asn 1791-1794）gaattc-gaactc
Sense primer: CTTAAGAACTCAAACAAAAGAAGCG
Anti-sense primer: TTTGTTTGAGTTCTTAAGCTCCTCC
Source
The phenylpyruvate decarboxylase from S. cerevisiae (Aro10, NP 010668.3).
References
Wang, P., Yang, X., Lin, B., Huang, J., & Tao, Y. (2017). Cofactor self-sufficient whole-cell biocatalysts for the production of 2-phenylethanol. Metabolic Engineering, 44(August), 143–149.