Team:Lethbridge/Improve
Methionine Synthetase (MetRS)
Original part: BBa_K567015
Submitted by: SJTU-BioX-Shanghai 2011
Designed by: You Wang
Rational behind improvements:
BBa_K567015 encodes for a truncated MetRS without the anticodon recognition domain. We have improved this part by including the entire coding sequence, which has been codon optimized for optimal expression in E. coli. For easy purification we have included a C- terminal hexahistidine tag with a serine glycine linker. We have also optimized MetRS to be overexpressed in BL21 DE3 gold cell by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015). The original part is also incompatible with biobrick standards. To address this we have removed all illegal cut sites from the construct.
Our Part:
T7 RNA Polymerase (T7 RNAP)
Original part: BBa_I2032
Submitted by: MIT
Designed by: Bartholomew Canton
Rational behind improvements: BBa_I2032 encodes exclusively for the coding region of T7 RNA polymerase. In order to improve it and incorporate it into our Next vivo system we have Codon optimized for use in E. coli and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized our T7 RNA polymerase to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).
Our Part:
Elongation Factor Thermo Stable (EF-Ts)
Original part: BBa_K1906000
Submitted by: XJTLU-China 2016
Designed by: Wenbo Xu
Rational behind improvements: BBa_K1906000 encodes exclusively for the coding region of EF-Ts. In order to improve it and incorporate it into our Next vivo system we have codon optimized for use in E. coli and attached a C-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Ts to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).
Our Part:
Elongation Factor Thermo Unstable (EF-Tu)
Original part:BBa_K1906004
Submitted by: XJTLU-China 2016
Designed by: Yuwei Han
Rational behind improvements: BBa_K1906004 encodes exclusively for the coding region of EF-Tu. In order to improve it and incorporate it into our Next vivo system we have codon optimized for use in E. coli and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Tu to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).
Our Part:
MS2 Coat Protein
Original part: BBa_K2109108
Submitted by: Lethbridge 2016
Designed by: Andy Hudson
Rational behind improvements: This year we utilized the MS2 coat protein (MS2BP) in our purification strategy of tRNA and ribosomes. As such, we expressed and purified MS2BP along with our tRNAPhe construct. From this we concluded that MS2BP expresses, binds to Nickel Sepharose affinity column, and binds the MS2 hairpin as designed.
Our Part:
