Team:Warwick/Results
Results
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Cloning
We have successfully cloned INP, BclA, and sZFs 2,10 & 14 individually into our construct gene, using the restriction sites we included in the design. Results were confirmed by sequencing colonies picked from antibiotic selection plates.
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Experiment 1: Testing the binding of specifically designed DNA strands to glass slides
We showed that we could bind aminated DNA to slides using a fluorescein-labelled oligonucleotide. Two slides were used: a GOPTS-treated slide that should bind aminated DNA, and a control slide that was cleaned like the GOPTS slide, but was not treated with the GOPTS solution, and should not bind DNA (aminated or otherwise). Our experiment showed fluorescence only on the treated slide, showing that our DNA-immobilisation technique was valid.
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Experiment 2: Testing the expression of zinc finger proteins on the surface of E. coli cells upon induction with IPTG
Our objective was to prove that the modified protein was being expressed on the surface of the plasmid transformed cells.
Zif268
- A: Wild type DH5α-Z1 E. coli cells
- B: Uninduced Lpp_OmpA-Zif268 transformed cells
- C: Induced Lpp_OmpA-Zif268 transformed cells
Upon comparison of image B and C,it is clear that the modified protein is being expressed upon induction with IPTG. However, the wild type cell shows fluorescence as well, suggesting that there is an error with our method of washing the cells after the fluorescent antibodies have been applied. The affected cells would fluoresce regardless of surface zinc finger expression.