Team:Austin Texas/photocage

From 2014.igem.org

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(Introduction)
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We recreated a light-activatable T7 RNA polymerase (RNAP) for the spatio-temporal control of protein expression. The light-activatable T7 RNAP was created by mutating a tyrosine codon at position 639 of a domain crucial for the polymerization of RNA during transcription.  Y639 was mutated to an amber codon, allowing us to incorporate a ncAA at this position.  We used ortho-nitrobenzyl tyrosine (ONBY), which is a "photocaged" ncAA (Figure 1).  Thus, if our synthetase/tRNA pair works, position 639 should contain ONBY in place of tyrosine.  This work is essentially a recapitulation of earlier work done by [reference authors/paper].
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We recreated a light-activatable T7 RNA polymerase (RNAP) for the spatio-temporal control of protein expression. The light-activatable T7 RNAP was created by mutating a tyrosine codon at position 639 of a domain crucial for the polymerization of RNA during transcription.  Y639 was mutated to an amber codon, allowing us to incorporate a ncAA at this position.  We used ortho-nitrobenzyl tyrosine (ONBY), which is a "photocaged" ncAA (Figure 1).  Thus, if our synthetase/tRNA pair works, position 639 should contain ONBY in place of tyrosine.  This work is essentially a recapitulation of earlier work done by [Chou et al. 2010].
    
    

Revision as of 00:09, 15 October 2014