Team:British Columbia/ProjectChassis

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<h3>Results</h3>
<h3>Results</h3>
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For the construction of the T7 replication machinery we cloned the DNAP, RNAP, ssDNA-BP and the primase into the pSB1C3 vector. All genes are under the control of the pBAD promoter to allow for titratable induction with arabinose. To test the expression of the T7 genes we transformed the plasmids into DH5α cells. Cloning of the primase, unfortunately, was troublesome and our expression experiments showed that the primase was defective. The DNAP and ssDNA-BP expressed to high levels (Fig 2). RNAP expression was not detectable on a SDS-Gel. We are planning to assemble the genes together with promoter and terminator on the pSB1C3 plasmids.  
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For the construction of the T7 replication machinery, we cloned the DNAP, RNAP, ssDNA-BP and the primase into the pSB1C3 vector. All genes are under the control of the pBAD promoter to allow for titratable induction with arabinose. To test the expression of the T7 genes we transformed the plasmids into DH5α cells. Cloning of the primase, unfortunately, was troublesome and our expression experiments showed that the primase was defective. The DNAP and ssDNA-BP expressed to high levels (Fig 2). RNAP expression was not detectable on a SDS-Gel. We are planning to assemble the genes together with promoter and terminator on the pSB1C3 plasmids.  
The JH-pSB2K3 with its T7 origin sequence will serve as vector for the GOI and will be transformed into cells with the T7 replication machinery.
The JH-pSB2K3 with its T7 origin sequence will serve as vector for the GOI and will be transformed into cells with the T7 replication machinery.

Revision as of 04:00, 18 October 2014

2014 UBC iGEM

© 2014 UBC iGEM