Experimental results

The experimental results for the Self Destructive Plasmid are detailed below. In the process of constructing it, we deviated from the cloning strategy after proving that it didn't work as expected.

I-SceI restriction site

Construction of the I-SceI homing endonuclease restriction site was done by primer synthesis. Primers were annealed and ligated into a pSB1AK3 backbone (3189bp without the insert). Together with the 30bp recognition sequence (shown below) this gave a 3219 bp part. The sites where cleavage of the DNA backbone occurs are indicated with ^.

5' A G T T A C G C T A G G G A T A A^C A G G G T A A T A T A G 3'
3' T C A A T G C G A T C C C^T A T T G T C C C A T T A T A T C 5'

I-SceI homing endonuclease

Integration

Experimental results

Experimental results

'Experimental results'

Experimental results

Experimental results

Experimental results

'Experimental results'

Experimental results

Experimental results

Experimental results

'Experimental results'

Experimental results