Part:BBa_K4288005
cadA promoter-signal peptide of RpmG-ferulyol esterase
cadA promoter-signal peptide of RpmG-ferulyol esterase
Characterization by Fujian_United
BBa_K4288005
Name: cadA promoter-signal peptide of RpmG-feruloyl esterase
Base Pairs: 1128 bp
Origin: Bacillus subtilis, genome
Properties: response to the cadmium concentration
Usage and biology
The biosensor used to monitor the cadmium and zinc in the environment.
Construct design
The reporter feruloyl esterase gene controlled under the cad promoter which monitors the cadmium and zine concentration of solution. Further, the reporter fused the signal peptide RpmG. The gene fragment was inserted into the pHY300PLK vector.
BBa_K4288000
Name: cadA promoter
Base Pairs: 159 bp
Origin: Bacillus subtilis, 50S ribosomal protein
Properties: signal peptide
Usage and biology
The cadmium resistance in bacteria using Cad A system. in high concentration of cadmium, the cad A system uptake the cadmium by ATP-dependent pump.
BBa_K4288002
Name: signal peptide of RpmG
Base Pairs: 87 bp
Origin: Bacillus subtilis, 50S ribosomal protein
Properties: signal peptide
Usage and biology
BBa_K4288002 is a coding sequence of RpmG, which is an amino acid sequence of 1-29. we used it as signal peptide.
BBa_K4288001
Name: Feruloyl esterase
Base Pairs: 870 bp
Origin: E.coli
Properties: hydrolysis the ester bands
Usage and biology
Feruloyl esterase (FAE) produced from bacteria, fungi and plant. The enzyme mainly functions as hydrolysis the ester bands. based on amino acids sequence alignment and substrate specificities, the feruloyl esterase classified into four types, including A, B, C, and D. The type D of feruloyl esterase acts as xylanase, which is the hemicellulolytic enzyme. thus, the feruloyl esterase was used as additive to improve the bioconversion and utilization of the agricultural stuff.
Improvement of the existing parts
Our composite part BBa_K4288005 was improved based on the existing part BBa_K174015 submitted by iGEM09_Newcastle. In 2019, group iGEM_Gaston Day School combined this part with an RBS (B0036), RFP (E1010), and a double terminator (B0014), and has shown that this promoter is sensitive to cadmium down to 0.25M (p<0.01). Today, our team further improved the cadmium sensor by optimizing the sequence of cad promoter, fusing a new signal peptide of RpmG which helps protein secret outside the cell, and combining a report protein feruloyl esterase(FAE). Therefore, the new composite part BBa_K4288005 was developed.
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]
| None |