Team:SJTU-BioX-Shanghai/Project/Subproject1-1

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==Rare tRNA Amount==
==Rare tRNA Amount==
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===Design===
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In this part we have overexpressed rare tRNA<sup>Arg</sup>-AGG in the cell. The rare tRNA can recognize AGG codon on the mRNA.
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'''tRNA<sup>Arg</sup>-AGG''': tRNA<sup>Arg</sup>-AGG is over expressed under the control of trc promoter (induced by IPTG).
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This rare tRNA<sup>Arg</sup> can be charged with Arg by native '''Arginyl-tRNA Synthetase(ArgRS)''' in E.coli.
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[[image:11SJTU_Rare_03.jpg|center|tRNA Modulator]]
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'''RFP-6AGG''': we have inserted 6 AGG codons after the start codon ATG in the RFP gene.
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[[image:11SJTU_rare_00.jpg|500px|center]]
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===Action===
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When rare tRNA<sup>Arg</sup>-AGG is not over-expressed, RFP expression is hindered. When tRNA<sup>Arg</sup>-AGG is over-expressed, this tRNA can recognize the AGG codon on the mRNA so a large amount of RFP is produced.
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[[image:11SJTU_rare_01.jpg|500px|center]]
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===Result===
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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 tRNAArg-AGG.''']]
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[[File:11SJTU_rare_10.jpg|thumb|500px|center|''Fig 2'' Cells over-expressing tRNAArg-AGG emit bright red fluorescence as wild type RFP (first one from the left). Control (first one from the right) exhibits no red fluorescence.]]
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RFP has been largely produced in cells overexpressing tRNA<sup>Arg</sup>. No RFP can be observed in cells without rare tRNA overexpression.
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'''We have successfully controlled protein expression by controlling rare tRNA amount.'''

Revision as of 05:12, 28 October 2011

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    Rare tRNA Amount

    Design

    In this part we have overexpressed rare tRNAArg-AGG in the cell. The rare tRNA can recognize AGG codon on the mRNA.

    tRNAArg-AGG: 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.

    tRNA Modulator

    RFP-6AGG: we have inserted 6 AGG codons after the start codon ATG in the RFP gene.

    11SJTU rare 00.jpg

    Action

    When 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.

    11SJTU rare 01.jpg

    Result

    Fig 1. Confocal Microscope examining RFP expression. RFP has been largely produced in cells overexpressing tRNAArg-AGG.
    Fig 2 Cells over-expressing tRNAArg-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 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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