Difference between revisions of "Part:BBa K2443028"

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<partinfo>BBa_K2443028 short</partinfo>
 
<partinfo>BBa_K2443028 short</partinfo>
  
EF-Ts is a prokaryotic elongation factors. EF-Ts acts at the guanine nucleotide exchange factor for EF-Tu, catalyzing the release of guanosine diphosphate from EF-Tu. This enables EF-Tu to bind to a new guanosine triphosphate molecule, release EF-Ts, and go on to catalyze another aminoacyl tRNA addition. EF-Ts has been Codon optimized for use in <i>Escherichia coli</I> and contains a C-terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator. The expected size of EF-Ts is 32 kDa, corresponding to the band observed lane XX. Protein overexpression and purification was done according to the Lethbridge 2017 protocols and notebook found on the wiki (https://static.igem.org/mediawiki/2017/2/2c/Protein_Overexpression_.pdf and https://static.igem.org/mediawiki/2017/0/0d/October_.pdf).  
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EF-Ts is a prokaryotic elongation factors. EF-Ts acts at the guanine nucleotide exchange factor for EF-Tu, catalyzing the release of guanosine diphosphate from EF-Tu. This enables EF-Tu to bind to a new guanosine triphosphate molecule, release EF-Ts, and go on to catalyze another aminoacyl tRNA addition. EF-Ts has been Codon optimized for use in <i>Escherichia coli</I> and contains a C-terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator. The expected size of EF-Ts is 32 kDa, corresponding to the band observed lane 9. Protein overexpression and purification was done according to the Lethbridge 2017 protocols and notebook found on the wiki (https://static.igem.org/mediawiki/2017/2/2c/Protein_Overexpression_.pdf and https://static.igem.org/mediawiki/2017/0/0d/October_.pdf).  
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https://static.igem.org/mediawiki/parts/c/c8/Lethbridge_oct_5_gel_3.png
  
 
<h1> Improved Part </h1>
 
<h1> Improved Part </h1>

Revision as of 20:32, 1 November 2017


EF-Ts optimized for expression in E. coli

EF-Ts is a prokaryotic elongation factors. EF-Ts acts at the guanine nucleotide exchange factor for EF-Tu, catalyzing the release of guanosine diphosphate from EF-Tu. This enables EF-Tu to bind to a new guanosine triphosphate molecule, release EF-Ts, and go on to catalyze another aminoacyl tRNA addition. EF-Ts has been Codon optimized for use in Escherichia coli and contains a C-terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator. The expected size of EF-Ts is 32 kDa, corresponding to the band observed lane 9. Protein overexpression and purification was done according to the Lethbridge 2017 protocols and notebook found on the wiki (https://static.igem.org/mediawiki/2017/2/2c/Protein_Overexpression_.pdf and https://static.igem.org/mediawiki/2017/0/0d/October_.pdf).

Lethbridge_oct_5_gel_3.png

Improved Part

Original part:BBa_K1906000 submitted by XJTLU-China 2016

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-Gold (DE3) cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 65
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]