Team:TU-Delft/receptordesign
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<h3>What?</h3> | <h3>What?</h3> | ||
Protocol for making protein chimeras with a rat G protein coupled receptor (RI7) and Your Favorite Receptor. The order of the DNA sequence looks like this: RI7-[Your Favorite Receptor]-RI7 | Protocol for making protein chimeras with a rat G protein coupled receptor (RI7) and Your Favorite Receptor. The order of the DNA sequence looks like this: RI7-[Your Favorite Receptor]-RI7 | ||
- | <h3>Why?</h3 | + | <h3>Why?</h3> |
One of the requirements for a working GPCR is that the receptor should be localized into the outside membrane of yeast cell. By replacing the N-terminal part of Your Favorite Receptor by the N-terminal ends of a receptor that is known to be localized into the outside membrane of Saccharomyces cerevisiae (R17), Your Favorite Receptor (YFR) will also be localized into the membrane. The C-terminal part of a GPCR is the alpha subunit binding region. If this is replaced by the RI7 regions a higher affinity with the alpha subunit can be reached [1]. | One of the requirements for a working GPCR is that the receptor should be localized into the outside membrane of yeast cell. By replacing the N-terminal part of Your Favorite Receptor by the N-terminal ends of a receptor that is known to be localized into the outside membrane of Saccharomyces cerevisiae (R17), Your Favorite Receptor (YFR) will also be localized into the membrane. The C-terminal part of a GPCR is the alpha subunit binding region. If this is replaced by the RI7 regions a higher affinity with the alpha subunit can be reached [1]. | ||
- | <h3>How?</h3 | + | <h3>How?</h3> |
With this step-by-step protocol we guide you trough all the in silico designing steps. After this the DNA can be transformed in yeast and you have your own olfactory yeast! | With this step-by-step protocol we guide you trough all the in silico designing steps. After this the DNA can be transformed in yeast and you have your own olfactory yeast! | ||
<p><br> | <p><br> |
Revision as of 07:28, 25 October 2012
Content
Chimeric receptor design: What, Why and HowIn silico protocol
Example
Chimeric receptor design: What, Why and How
What?
Protocol for making protein chimeras with a rat G protein coupled receptor (RI7) and Your Favorite Receptor. The order of the DNA sequence looks like this: RI7-[Your Favorite Receptor]-RI7Why?
One of the requirements for a working GPCR is that the receptor should be localized into the outside membrane of yeast cell. By replacing the N-terminal part of Your Favorite Receptor by the N-terminal ends of a receptor that is known to be localized into the outside membrane of Saccharomyces cerevisiae (R17), Your Favorite Receptor (YFR) will also be localized into the membrane. The C-terminal part of a GPCR is the alpha subunit binding region. If this is replaced by the RI7 regions a higher affinity with the alpha subunit can be reached [1].How?
With this step-by-step protocol we guide you trough all the in silico designing steps. After this the DNA can be transformed in yeast and you have your own olfactory yeast!