Team:IBB Pune/protein-based-signalling
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- | =The Complete Turing Machine | + | {{Team:IBB_Pune/header}} |
- | + | {{Team:IBB_Pune/menu}}<br><br> | |
+ | <p><span style="font-weight:bold; font-size:200%; color:#0000cc;">The Complete Turing Machine</span></p> | ||
[[Image: BBa_K233325.JPG|center|900px]] | [[Image: BBa_K233325.JPG|center|900px]] | ||
- | The above image is a representation of the complete turing machine including inter state regulation. This model is true to Alan Turing's vision of multi-mutually exclusive states and the states being regulated by one another through outputs. | + | The above image is a representation of the complete turing machine including inter state regulation. This model is true to Alan Turing's vision of multi-mutually exclusive states and the states being regulated by one another through outputs.<br> |
- | + | <p><span style="font-weight:bold; font-size:150%; color:#FF6600;">Working of the Construct</span></p> | |
- | + | [[Image:Complicated.png|center|800px]] | |
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In the above construct, there are three independent modules that function | In the above construct, there are three independent modules that function | ||
* A constitutively working module which produces LuxR protein and LacI and Cassette 1 (encodes ogr and activates state 'B') | * A constitutively working module which produces LuxR protein and LacI and Cassette 1 (encodes ogr and activates state 'B') | ||
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Cassette 2 also encodes for the phage lambda cI repressor protein which represses cassette 1. Thus we have a system in which the two states are mutually exclusive. In state 'B', when the machine encounters a '1', it keeps the '1' unchanged and transits to the next cell. | Cassette 2 also encodes for the phage lambda cI repressor protein which represses cassette 1. Thus we have a system in which the two states are mutually exclusive. In state 'B', when the machine encounters a '1', it keeps the '1' unchanged and transits to the next cell. | ||
- | Now in this state, when the machine encounters a '0' the AND gate obtains both its inputs viz. ogr and lactose which results in the production of Homoserine Lactone Synthase (enzyme producing AHL). This results in the conversion of the '0' to a '1'. Therefore this construct behaves like a unary adder as per Turing's specification. | + | Now in this state, when the machine encounters a '0' the AND gate obtains both its inputs viz. ogr and lactose which results in the production of Homoserine Lactone Synthase (enzyme producing AHL). This results in the conversion of the '0' to a '1'. Therefore this construct behaves like a unary adder as per Turing's specification.<br> |
+ | <p><span style="font-weight:bold; font-size:150%; color:#FF6600;">Problems Associated with such an Approach</span></p> | ||
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*At 6.5kb the construct too darn BIG! | *At 6.5kb the construct too darn BIG! | ||
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*It poses major complications with respect to quality control, sequencing and data validation. | *It poses major complications with respect to quality control, sequencing and data validation. | ||
- | =Modular Systems | + | One of the methods of solving these problems is to [https://2009.igem.org/Team:IBB_Pune/construct simplify] the construct.<br> |
+ | <p><span style="font-weight:bold; font-size:150%; color:#FF6600;">Modular Systems</span></p> | ||
+ | |||
We decided to use a modular approach to tackle the problems enlisted above. | We decided to use a modular approach to tackle the problems enlisted above. | ||
We therefore decided to divide the complicated construct into three separate interdependent strains which together combine to form the Unary Adder. | We therefore decided to divide the complicated construct into three separate interdependent strains which together combine to form the Unary Adder. | ||
- | + | [[Image:Asdasdasd.png|center|968px]] | |
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In this system the first module gets activated in the presence of AHL ( '1' ) and produces ogr attached to an export tag (YcdB or TorA) this results in the export of ogr protein which acts as an input for the second module. | In this system the first module gets activated in the presence of AHL ( '1' ) and produces ogr attached to an export tag (YcdB or TorA) this results in the export of ogr protein which acts as an input for the second module. | ||
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This functions as a unary adder | This functions as a unary adder | ||
- | + | 9 | |
- | =Proof-of-concept | + | <br><p><span style="font-weight:bold; font-size:150%; color:#FF6600;">Proof-of-concept</span></p> |
As a proof of concept of the export-based signalling machinery, we attempted to test the working of the export tags by fusing them upstream of Green Fluorescent Protein. | As a proof of concept of the export-based signalling machinery, we attempted to test the working of the export tags by fusing them upstream of Green Fluorescent Protein. | ||
<html> | <html> | ||
<p><a href="https://2009.igem.org/SNOWDRIFT/Proof_of_concept | <p><a href="https://2009.igem.org/SNOWDRIFT/Proof_of_concept | ||
- | "><span style="font-weight:bold; font-size:150%; color:# | + | "><span style="font-weight:bold; font-size:150%; color:#0000cc;"><u>NEXT</u></span></a></p> |
</html> | </html> |
Latest revision as of 02:39, 22 October 2009