Team:Tsinghua/Experiment

From 2009.igem.org

(Difference between revisions)
(Synthesis of the Gene Therapy Production System)
(Project 1)
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Specifically, we choose two molecular cloning vectors to encode two sections of the gene therapy vector genome, pET28a and pACYCDuet1.
Specifically, we choose two molecular cloning vectors to encode two sections of the gene therapy vector genome, pET28a and pACYCDuet1.
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[[Image:pET28a.jpg|pET28a]]
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[[Image:pACYCDuet1.jpg|pACYCDuet1]]
===Top-Down Approach===
===Top-Down Approach===

Revision as of 16:20, 15 August 2009

Contents

Project 1

Synthesis of the Therapeutic DNA

In this part, we aims at constructing a molecular cloning vector with a cos site which enables it to be packaged into the gene therapy vector- in other words, to construct a cosmid. In order to detect the expression of the therapeutic DNA, we construct this cosmid based on the scanfold of Parts-J61031 encoding a RFP-expressing segment.

This cosmid mainly consists of the following segments:

1) origin of replication: we insert O gene and P gene from bacteriophage lambda into J61031, which are resoonsible for the late phase replication and package of the wild type circular bacteriophage lambda genome

2) cos site: necessary for the specific package of the Therapeutic DNA into the gene therapy vector.

3) RFP-expressing segment: originally integrated into the plamid of J61031.

cosmidcostruct

Synthesis of the Gene Therapy Production System

Bottom-Up Approach

In the bottom-up approach, we amplify the biobricks from both the bacteriophage lambda and the adenovirus genome and incorporate them with a given order into molecular cloning vector(s).

Specifically, we choose two molecular cloning vectors to encode two sections of the gene therapy vector genome, pET28a and pACYCDuet1.

pET28a pACYCDuet1

Top-Down Approach

Synthesis of the Targeted Biobrick

Production of the targeted gene therapy vector

Functioning of the targeted gene therapy vector

Project 2

Project 1 and Project 2