From 2014.igem.org
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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 the O-helix ('''figure??? of helix?'''), which is 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]. | + | 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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Revision as of 23:47, 14 October 2014