Difference between revisions of "Part:BBa K2683017"
 
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<partinfo>BBa_K2683017 short</partinfo>
 
<partinfo>BBa_K2683017 short</partinfo>
  
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Release factor 2 (RF2) is a protein that causes termination of translation by recognizing the stop codon in an mRNA sequence. This part was originally designed by Lethbridge iGEM team 2017(http://2017.igem.org/Team:Lethbridge) The original composite part can be found here: https://parts.igem.org/Part:BBa_K2443030 RF2 has been Codon optimized for use in Escherichia coli and contains a N-Terminal hexahistidine tag. It is under the regulation of T7 Promoter, RBS and double terminator.
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Release Factor 2 (RF2) is a protein that causes termination of translation by recognizing the stop codon in an mRNA sequence. This part was originally designed by Lethbridge iGEM team 2017(http://2017.igem.org/Team:Lethbridge) The original composite part can be found here: https://parts.igem.org/Part:BBa_K2443030 RF2 has been Codon optimized for use in Escherichia coli and contains a N-Terminal hexahistidine tag. It is under the regulation of T7 Promoter, RBS and double terminator.
  
 
We have improved the part by successfully inserting it into the pSB1C3 standard plasmid.  
 
We have improved the part by successfully inserting it into the pSB1C3 standard plasmid.  
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https://static.igem.org/mediawiki/2018/5/50/T--Lethbridge--RF1RF2.PNG
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Figure 1- The overexpression of Release Factor 2 in pSB1C3 as run on a 12% SDS-PAGE gel. Samples taken correspond to point of induction and  samples taken over the next three hours. The protein is expected at about 43kDa.
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https://static.igem.org/mediawiki/2018/f/f8/T--Lethbridge--RF2pur.PNG
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Figure 2- 12% SDS-PAGE showing the nickel affinity chromatography purification of Release Factor 2. Elutions 1-5 show the purified proteins at the 50kDa range.
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<!-- Add more about the biology of this part here.
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===Usage and Biology===
 
===Usage and Biology===
  

Latest revision as of 03:17, 17 October 2018


Release Factor 2 in pSB1C3

Release Factor 2 (RF2) is a protein that causes termination of translation by recognizing the stop codon in an mRNA sequence. This part was originally designed by Lethbridge iGEM team 2017(http://2017.igem.org/Team:Lethbridge) The original composite part can be found here: https://parts.igem.org/Part:BBa_K2443030 RF2 has been Codon optimized for use in Escherichia coli and contains a N-Terminal hexahistidine tag. It is under the regulation of T7 Promoter, RBS and double terminator.

We have improved the part by successfully inserting it into the pSB1C3 standard plasmid.

T--Lethbridge--RF1RF2.PNG

Figure 1- The overexpression of Release Factor 2 in pSB1C3 as run on a 12% SDS-PAGE gel. Samples taken correspond to point of induction and samples taken over the next three hours. The protein is expected at about 43kDa.

T--Lethbridge--RF2pur.PNG

Figure 2- 12% SDS-PAGE showing the nickel affinity chromatography purification of Release Factor 2. Elutions 1-5 show the purified proteins at the 50kDa range.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 659
    Illegal BglII site found at 1116
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]