Part:BBa_K127004:Experience

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••••• iGEM12_OUC-China

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We improved this part,providing the improved partK737034 with new function. Fusion of galK to the 2nd codon of GFP, gives this constitutive GFP generator ability to monitor the Spot42 level in cytosol. galK is the leader sequence of galactokinase (GalK),and Spot42 small RNA controls the synthesis of the galactokinase (GalK) in response to the availability of glucose in the environment,by blocking the recognition of antiSD motif to RBS.The repression is modest(about 2.6 fold),maybe there is no subsequent degradation caused by the major degradase,RNaesE and RNaseIII. Experimental data: We ligated this constitute galk::GFP generator to spot42 generator controlled by aTc concentration, tested the binding affinity between spot42 and galK::mRNA indirectly through monitoring fluorescence. Fig. 1 This is aTc induction experiment of K737039. Production of spot42 is induced by aTc. The translation of galK::GFP mRNA will be inhibited by spot42 in our expectation. And experimental results show that aTc induction works well for inhibition of galK::GFP mRNA expression which is represented with decreasing RFU. As we have expected,the spot42 repression efficiency on galK is modest NOTE:The test is perform on a medium copy number plasmid,J61002.