Team:SJTU-BioX-Shanghai/Project/Subproject1-1
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[[File:11SJTU_rare_02.png|thumb|500px|center|''Fig 1''. Confocal Microscope examining RFP expression. '''RFP has been largely produced in cells overexpressing tRNA(Arg)-AGG.''']] | [[File:11SJTU_rare_02.png|thumb|500px|center|''Fig 1''. Confocal Microscope examining RFP expression. '''RFP has been largely produced in cells overexpressing tRNA(Arg)-AGG.''']] | ||
- | [[File:11SJTU_rare_10.jpg|thumb|500px|center|''Fig 2'' Cells over-expressing tRNA | + | [[File:11SJTU_rare_10.jpg|thumb|500px|center|''Fig 2'' Cells over-expressing tRNA<sup>Arg</sup>-AGG emit bright red fluorescence as wild type RFP (first one from the left). Control (first one from the right) exhibits no red fluorescence.]] |
RFP has been largely produced in cells overexpressing tRNA<sup>Arg</sup>. No RFP can be observed in cells without rare tRNA overexpression. | RFP has been largely produced in cells overexpressing tRNA<sup>Arg</sup>. No RFP can be observed in cells without rare tRNA overexpression. |
Revision as of 18:02, 28 October 2011
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Rare tRNA AmountDesignIn this part we have overexpressed rare tRNAArg-AGG in the cell. The rare tRNA can recognize AGG codon on the mRNA. tRNAArg-AGG([http://partsregistry.org/wiki/index.php?title=Part:BBa_K567001 BBa_K567001]): tRNAArg-AGG is over expressed under the control of trc promoter (induced by IPTG). This rare tRNAArg can be charged with Arg by native Arginyl-tRNA Synthetase(ArgRS) in E.coli. RFP-6AGG([http://partsregistry.org/wiki/index.php?title=Part:BBa_K567017 BBa_K567017]): we have inserted 6 AGG codons after the start codon ATG in the RFP gene. ActionWhen rare tRNAArg-AGG is not over-expressed, RFP expression is hindered. When tRNAArg-AGG is over-expressed, this tRNA can recognize the AGG codon on the mRNA so a large amount of RFP is produced.
ResultRFP has been largely produced in cells overexpressing tRNAArg. No RFP can be observed in cells without rare tRNA overexpression. We have successfully controlled protein expression by controlling rare tRNA amount.
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