Genome shuffling

BioBrick

Hbsu-(x)fp BioBrick

Purpose of BioBrick

Tag the bacterial chromosome with a fluorescent protein, in order to observe genetic recombination between two bacteria by genome shuffling.

Introduction to Project

B. subtilis L-forms will be squeezed together using microfluidics in the presence of PEG (polyethylene glycol, which facilitates cellular fusion). Fusion will result in the genomes coming into close proximity. The L-forms being fused are from the same strain; therefore their genomes will be almost identical. This will allow homologous recombination, in which similar sections of DNA are ‘swapped’ between genomes, to take place. Subsequent splitting of the fusion product will give two new cells, neither of which is identical to their ‘parents’.

We are exploiting the diversity that naturally exists between members of the same species. Parallels can be drawn between this process and meiosis, and could even be viewed as making the bacteria procreate. Our new cells will be phenotypically distinct, in the same way that a child is different from either of its parents. Some of the cells produced will have enhanced function of the trait of interest. This can be assayed for, these cells isolated, and the process repeated multiple times, giving us new and improved (for the phenotype of interest) cells. In theory, different strains, or even species, of bacteria could be fused and exhibit shuffling, increasing variation further.

Purpose

Over the past eight years hundreds of iGEM teams have dreamt up exciting and innovative ways of harnessing Synthetic Biology. However only a few iGEM projects have developed into further research. Indeed, despite the thousands of hours devoted to research, the field of Synthetic Biology has not made a viable product.

Synthetic Biology is a relatively new subject, and like similar, more established areas, such as Genetic Engineering it has not yet delivered on its potential.

It would be disingenuous to downplay the important research happening around the world in these fields. However it is undeniable that a gap exists between what we want to achieve and what is currently achievable. Whilst this lack of progress can be attributed to both the complexity of biological systems involved and a public suspicious of genetic modification, there is also the problem of producing a viable, high quality yield of the desired product.

Processes in biology can be refined and improved by evolution, but in nature this occurs very slowly. We propose using L-forms to artificially evolve B. subtilis using genome shuffling. This technique increases the rate of evolution, allowing the improvement of any biological system from theoretically thousands of years, to months.