Difference between revisions of "Team:Heidelberg/Optogenetics"

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The generation of proteins with radically altered or highly specific new activities is a very challenging task in the field of directed evolution. Modulation of selection stringency is indispensable for the optimization of proteins, which in their native form exhibit a lack of activity for the required action. To further extent the options of PACE/PREDCEL based protein optimization and to evolve new activities from an initially inactive gene without the use of intermediate evolutionary steps, we provide an optogenetic modulator of selection stringency. The variation of stringency is based on the blue light-dependent transcription factor EL222 and a bidirectional promoter system that can induce or repress the expression of geneIII upon blue light irradiation in a non-toxic, rapidly delivered, and reversible manner.
 
The generation of proteins with radically altered or highly specific new activities is a very challenging task in the field of directed evolution. Modulation of selection stringency is indispensable for the optimization of proteins, which in their native form exhibit a lack of activity for the required action. To further extent the options of PACE/PREDCEL based protein optimization and to evolve new activities from an initially inactive gene without the use of intermediate evolutionary steps, we provide an optogenetic modulator of selection stringency. The variation of stringency is based on the blue light-dependent transcription factor EL222 and a bidirectional promoter system that can induce or repress the expression of geneIII upon blue light irradiation in a non-toxic, rapidly delivered, and reversible manner.
 
In this subproject, we demonstrate a blue light-dependent increase in phage propagation using the pBLind-geneIII cassette. This part of our toolbox offers the scientific community an easy adaptation of the selection stringency in PACE and PREDCEL experiments and paves the way for a bright future in protein optimization.  
 
In this subproject, we demonstrate a blue light-dependent increase in phage propagation using the pBLind-geneIII cassette. This part of our toolbox offers the scientific community an easy adaptation of the selection stringency in PACE and PREDCEL experiments and paves the way for a bright future in protein optimization.  

Revision as of 19:06, 31 October 2017


Optogenetics
Modulator of selection stringency
The generation of proteins with radically altered or highly specific new activities is a very challenging task in the field of directed evolution. Modulation of selection stringency is indispensable for the optimization of proteins, which in their native form exhibit a lack of activity for the required action. To further extent the options of PACE/PREDCEL based protein optimization and to evolve new activities from an initially inactive gene without the use of intermediate evolutionary steps, we provide an optogenetic modulator of selection stringency. The variation of stringency is based on the blue light-dependent transcription factor EL222 and a bidirectional promoter system that can induce or repress the expression of geneIII upon blue light irradiation in a non-toxic, rapidly delivered, and reversible manner. In this subproject, we demonstrate a blue light-dependent increase in phage propagation using the pBLind-geneIII cassette. This part of our toolbox offers the scientific community an easy adaptation of the selection stringency in PACE and PREDCEL experiments and paves the way for a bright future in protein optimization.
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