Team:Birkbeck/Conclusion

• 
• Team
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  • Attributions
• Research
  • Overview
  • Background
  • Experiments
  • Measurements
  • Modelling
  • Results
  • Conclusion
  • Safety
• BioBricks
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  • Human Practices
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• InterLab
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Conclusion

Signal Detection

As mentioned in the discussion, there has been success in the transduction of Mycobacterium cells using mycobacteriophages. These protocols require growing up cells, exposing cells to bacteriophages & conducting a plaque assay. The preliminary results from our study show that fluorescence can be detected from as early as 20-60 minutes of E. coli growth. This would greatly reduce the time frame of pathogen detection. The results of the interlab study also show that the protocol is adaptable to accommodate different laboratory environments.

Biobricks For Mutagenisis

The stf gene was successfully reconstructed with orf-401 & orf-314. No mutagenesis work was carried out on stf. However, the biobrick is now available for mutagenesis work & future testing for changes in binding specificity.

The TFA acts as a molecular chaperone STF. The cloning of tfa will permit future mutagenesis work, which may facilitate the assembly of mutant STF if problems arise when mutations are introduced to stf. tfa was only cloned into the circuit & not made into a single biobrick. The tfa gene was cloned into the circuit where it is repressed by TetR. The addition of anhydroustetracycline will alleviate the repression and therefore allow optimization of tfa expression to give the largest yield of recombinant virions possible.

Biobricks for the Lytic/Lysogenic Cycle Control

For the biosynthesis of bacteriophage virions for our product, it is essential that the lytic & lysogenic cycles are tightly regulated in order to maximise the number of virions produced. The CI & Cro genes were successfully constructed into a circuit. With CI being constituetively expressed, lysogeny can be maintained for bacterial load to be increased in order to maximise the number of bacteriophage virions to be produced. The T7-RNAP is controlled under pBAD promoter which in turn stimulates the expression of Cro. This control can allow optimization of when to stimulate the switch from lysogeny to lytic cycle to maximise viral titre in the lysate.