Difference between revisions of "Part:BBa K2683017"
 
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https://static.igem.org/mediawiki/2018/5/50/T--Lethbridge--RF1RF2.PNG
 
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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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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===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]