Team:Kyoto/GSDD/Goals

From 2009.igem.org

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Our real goal is to suggest some possibility of realization of our ultimate goal.  
Our real goal is to suggest some possibility of realization of our ultimate goal.  
We will challenge to make a “prototype” device of our ultimate goal. Initially, we had planned to construct the designed two vectors (Timer vector and Bomb vector) and analyze those functions. However, after considering the span of experimental period, we changed our plan and decided to concentrate on the construction and functional analysis of Timer vector system (see details in Mechanism page).  
We will challenge to make a “prototype” device of our ultimate goal. Initially, we had planned to construct the designed two vectors (Timer vector and Bomb vector) and analyze those functions. However, after considering the span of experimental period, we changed our plan and decided to concentrate on the construction and functional analysis of Timer vector system (see details in Mechanism page).  
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We designed Timer vector to constitutively express EGFP while keeping the repetitive sequences of LacI binding site to be stabilize the DNA. In addition, we designed constitutive LacI expression vector (Totally two vectors). After the plasmid construction, we will move on to the functional measurement of Timer vector system; whether LacI binds to the both ends of repeated sequences and stabilized the linear Timer vector sufficiently and whether Timer vector can behave like natural linear chromosome and roughly count the replication times. (See below box and subgoals).  
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We designed Timer vector to constitutively express EGFP while keeping the repetitive sequences of LacI binding site to be stabilize the DNA. In addition, we designed constitutive LacI expression vector (totally two vectors). After the plasmid construction, we will move on to the functional measurement of Timer vector system; whether LacI binds to the both ends of repeated sequences and stabilized the linear Timer vector sufficiently and whether Timer vector can behave like natural linear chromosome and roughly count the replication times. (See below box and subgoals.)
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[[Image:figure(realgoal).png|498px|center|Fig.2]]
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[[Image:figure(realgoal).png|498px|center|Fig.1]]
===Subgoals===
===Subgoals===

Revision as of 18:48, 21 October 2009

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Goals

Ultimate Goal

Our ultimate goal is to make a device which enables us to control exact cell’s life time (or death time) depending on the number of cell division times.

Real Goal

Our real goal is to suggest some possibility of realization of our ultimate goal. We will challenge to make a “prototype” device of our ultimate goal. Initially, we had planned to construct the designed two vectors (Timer vector and Bomb vector) and analyze those functions. However, after considering the span of experimental period, we changed our plan and decided to concentrate on the construction and functional analysis of Timer vector system (see details in Mechanism page). We designed Timer vector to constitutively express EGFP while keeping the repetitive sequences of LacI binding site to be stabilize the DNA. In addition, we designed constitutive LacI expression vector (totally two vectors). After the plasmid construction, we will move on to the functional measurement of Timer vector system; whether LacI binds to the both ends of repeated sequences and stabilized the linear Timer vector sufficiently and whether Timer vector can behave like natural linear chromosome and roughly count the replication times. (See below box and subgoals.)

Fig.1

Subgoals

To attain the real goal, we set up the four subgoals in this project.


Subgoal-1

Simple construction of the repetitive sequences (TIMER) that contain multiple protein binding sites by Microgene Polymerization Reaction (MPR).

Subgoal-2

Construction of Timer vector that contains LacI generator and TIMER.

Subgoal-3

Construction of the constitutive LacI expression vector

Subgoal-4

Analysis of the constructed TIMER vector in yeast