Team:Austin Texas/kit

From 2014.igem.org

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(Background)
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Complications arise when the genetic code is recoded. In a normal bacterium, release factor RF1 is responsible for terminating translation when the ribosome reaches the amber stop codon. To avoid termination at a UAG amber codon, a strain of ''E. coli'' was engineered by the Church and Isaacs groups using MAGE and CAGE ('''ref''') to remove all of the amber codons from the genome and knock out the RF1 gene. The resulting strain, called "amberless" ''E. coli'', has its amber codon free to code for any ncAA. During translation, a synthetase with mutations that allow the acceptance of a different amino acid than the wild type charges that ncAA onto a tRNA with the amber codon's anticodon, CUA, when both are present in the cell.
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Complications arise when the genetic code is recoded. In a normal bacterium, release factor RF1 is responsible for terminating translation when the ribosome reaches the amber stop codon. To avoid termination at a UAG amber codon, a strain of ''E. coli'' was engineered by the Church and Isaacs groups using MAGE and CAGE to remove all of the amber codons from the genome and knock out the RF1 gene. The resulting strain, called "amberless" ''E. coli'', has its amber codon free to code for any ncAA. During translation, a synthetase with mutations that allow the acceptance of a different amino acid than the wild type charges that ncAA onto a tRNA with the amber codon's anticodon, CUA, when both are present in the cell.
=Experimental Design and Method=
=Experimental Design and Method=

Revision as of 23:21, 16 October 2014