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===Usage=== | ===Usage=== | ||
Latest revision as of 00:12, 22 October 2021
T7_Promoter-RBS-TnaA-T7_Terminator
A part for TnaA expression in the preliminary experiment.
Biology
TnaA cleaves 6-Br-Trp into 6-Br-indole, pyruvate and ammonia, so we can use this composite part to synthetic 6-Br-indole as the second step to product 6,6’-dibromo indigo.
Usage
We construct the composite part BBa_K3332052 and transformed it into E.coli DH5α& E. coli BL21 (DE3) to verify its expression.
Characterization
1.Identification
After plasmid construction, we performed PCR to certify and the resulting was shown down.
2.The proof of expression
We used IPTG to induce the expression at different times.
- Fig.1SDS-PAGE analysis of TnaA .Induction condition: LB media, OD600 0.6 ~ 0.8, IPTG 0.1 mM, 37 °C and 200 rpm.(1:negitive control 2: 9h 3:10.5h 4:12h 5:13.5h 6:15h 7:16.5h 8: 18h 9: 19.5h)
References
Kim, H. J. et al. Biosynthesis of indigo in Escherichia coli expressing self- sufficient CYP102A from Streptomyces cattleya. Dyes Pigments 140, 29–35 (2017).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]