"
Team:Tsinghua/Experiment1
From 2009.igem.org
Contents |
Synthesis of the Genome of GenSniper
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, pET-28a and pACYCDuet-1.
We incorporate the segment from Nu1 to B on bacteriophage lambda genome into pET-28a while clone the left segment from C to FII (C will be further engineered) into pACYCDuet-1. Before cotranformation with therapeutic DNA, we firstly cotranform the two plasmids encoding the GenSniper genome into BL21 DE3 and evaluate their function.
Top-Down Approach
In the top-down approach, the part of needed segment on the lambda genome will be firstly cutted by restriction enzyme BamHI and HindIII and then incorporated into pET-28a. The lefted segment will be further integrated downstream of the primary clone. Additionally, an expression module for Q protein will be contructed for induction of the expression of the GenSniper genome.
