Difference between revisions of "Part:BBa K3467007:Experience"

 
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CarH protein is from Thermus thermophilus. This protein has cobalamine (Vitamin B12), which has a photosensitive Co-C bond, as a prosthetic group. In the dark, CarH is a tetramer that binds strongly to CarO DNA operon and represses downstream protein translation. When excited by green light, the photosensitive Co-C bond breaks, this disrupts the tetramer structure, and then gene expression can start. From the research we know CarH maximum absorption spectrum is 525nm, we decided to apply green light control system to manipulate yeast dsRNA synthesis. When the yeast is incubated in dark condition, CarH will bind to dsRNA synthesis part which will block the dsRNA synthesis. When exposing to 525nm light environment, CarH tetramer photolysis which resulted in dsRNA synthesis expression.
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From the reference, CarH tetramer can bind with CarO operon and its induction fold can up to be 350. We inserted it in our yeast system with a TEF promoter on its upstream. We've already transformed it into yeast system and we'll do further research to test whether our CarH composite part can work well in the yeast system.
  
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__NOTOC__
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Chatelle, C. et al., A Green-Light-Responsive System for the Control of Transgene Expression in Mammalian and Plant Cells. ACS SYNTH BIOL 7 1349 (2018).
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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how you used this part and how it worked out.
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===Applications of BBa_K3467007===
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===User Reviews===
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<I>Username</I>
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Enter the review inofrmation here.
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Revision as of 17:00, 25 October 2020

CarH protein is from Thermus thermophilus. This protein has cobalamine (Vitamin B12), which has a photosensitive Co-C bond, as a prosthetic group. In the dark, CarH is a tetramer that binds strongly to CarO DNA operon and represses downstream protein translation. When excited by green light, the photosensitive Co-C bond breaks, this disrupts the tetramer structure, and then gene expression can start. From the research we know CarH maximum absorption spectrum is 525nm, we decided to apply green light control system to manipulate yeast dsRNA synthesis. When the yeast is incubated in dark condition, CarH will bind to dsRNA synthesis part which will block the dsRNA synthesis. When exposing to 525nm light environment, CarH tetramer photolysis which resulted in dsRNA synthesis expression. From the reference, CarH tetramer can bind with CarO operon and its induction fold can up to be 350. We inserted it in our yeast system with a TEF promoter on its upstream. We've already transformed it into yeast system and we'll do further research to test whether our CarH composite part can work well in the yeast system.

Chatelle, C. et al., A Green-Light-Responsive System for the Control of Transgene Expression in Mammalian and Plant Cells. ACS SYNTH BIOL 7 1349 (2018).