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images-8.jpegBioBricks and the standardisation of biological components

The use of standardised components is found in all fields of engineering, where it allows the benefits of abstraction, decoupling and insulation from unnecessary detail in the design of complex systems. The ability to construct new DNA sequences has far outpaced our capacity to design the genetic circuits that these might encode. Synthetic Biology promises simplified design using modular standardised DNA components, or BioBricks. 

 

Bacterial rainbow

 

The Cambridge iGEM2009 team has produced a series of gene expression cassettes that allow in vivo production of a spectrum of pigments in bacteria. More details can be found at http://2009.igem.org/Team:Cambridge/Project/Pigments

Carotenoids: The enzymes required for carotenoid production originally come from Pantoea ananatis, and were available in the registry. These were used to produce orange and red.
Melanin: The tyrosinase required for melanin production originally comes from Rhizobium etli and produces brown.
Violacein: The enzymes required for voilacein production came from Chromobacterium violaceum. The operon can be manipulated to produce violet, green, and blue.

   

Synthetic operon for violacein production

 

The Cambridge iGEM2009 team received sponsorship from DNA2.0 Inc., which allowed them to design and construct a synthetic operon for the biosynthesis of violacein. The operon is 7.5Kb in size, contains 5 genes, and has been submitted to the MIT Registry for Standard Parts in BioBrick format - Part BBa_K274002. Expression of the VioA-E genes results in conversion of L-Tyrosine to an intense violet pigment. Violacein is a hydrophobic compound, and is retained within cells.

   

New BioBrick encoding an improved fluorescent protein

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Green Fluorescent Protein (GFP) offers efficient and convenient means of visualising the dynamic process of gene expression and of obtaining readout of the current state of complex gene regulatory networks - features of major interest for synthetic biology.

Stefan Milde, working in the Haseloff Lab at Cambridge as part of iGEM2008  has constructed BioBrick versions of improved GFP variants and tested their properties (Parts:BBa I746908-I746919). He has compared two recently reported GFP variants to the mut3GFP variant in the Registry of Standard Biological Parts. The two GFP variants chosen were "superfolder GFP", developed and described by Pédelacq et al (2006), which was engineered for improved fluorescence in fusion proteins and P7 GFP ("superfast GFP") which was engineered by Fisher et al (2008) and selected on the basis of its very rapid folding in vitro. Read on for more...

   

The Registry of Standard Parts at MIT 

parts-icon
With BioBrick parts from Cambridge iGEM teams: iGEM2005, iGEM2006, iGEM2007, iGEM2008 and the Haseloff Lab and new Bacillus subtilis strains and key parts 
(http://partsregistry.org/) 
   
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