Difference between revisions of "Part:BBa K4711026"
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Revision as of 09:06, 10 October 2023
T7+alkB+alkG+T7+alkT+Spycatcher
Usage and Biology
Colony PCR results showed the appearance of the target band at around 4000 bp, indicating successful plasmid integration into Escherichia coli. SDS-PAGE analysis revealed the successful expression of the AlkB, AlkG, and AlkT proteins. Additionally, by setting a gradient of IPTG concentrations, it was observed that the protein expression was most efficient when induced with 0.5 mM IPTG, yielding the highest protein expression levels.
Fig 1 (A) Gene circuit (B) Colony PCR (C) SDS-PAGE M: Protein Marker 1: 0mM IPTG 2: 0.2mM IPTG 3: 0.5mM IPTG 4: 1mM IPTG. The protein bands from top to bottom are alkT (56KDa), alkB (45KDa), and alkG (18KDa).
Source
Pseudomonas oleovorans
Potential applications
References
[1]Qiaofei He, George N. Bennett, Ka-Yiu San, Hui Wu*. Biosynthesis of medium-chain ω-hydroxy fatty acids by AlkBGT of Pseudomonas putida GPo1 with native FadL in engineered Escherichia coli. Frontiers in Bioengineering and Biotechnology. 2019. 7:273.
[2] Staijen I E , Beilen J B V , Witholt B .Expression, stability and performance of the three-component alkane mono-oxygenase of Pseudomonas oleovorans in Escherichia coli[J].European journal of biochemistry, 2000, 267(7):1957-65.DOI:10.1046/j.1432-1327.2000.01196.x.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1860
Illegal NheI site found at 3534 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1634
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 167
Illegal NgoMIV site found at 1079
Illegal NgoMIV site found at 2554 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1966