Team:Freiburg/Project
From 2011.igem.org
Project overview
We aim to transform protein expression in bacterial systems into an elegant, fast and affordable process. By eliminating routine use of expensive materials, this novel assay will utilize sustainable laboratory equipment and widespread His-Tag technology.
We propose an expression system induced by blue, green and red light, combined with subsequent temperature controlled autolysis of E. coli. The purification of the his-tagged protein of interest will be accomplished by an adaptor protein of our own design which binds the His-Tag on one side and the surface of serological pipettes on the other. Two subsequent pipetting steps for washing and purification of the cell lysate will quickly elute the product.
Our system will provide expression and purification of Polymerase and Ligase, but will be easily expandable to any His-tagged protein needed by the modern molecular biologist.
About
Synthetic Biology can be an expensive discipline to conduct research in. It requires many kits, reagents and costly laboratory apparatus; all of which require an advanced infrastructure for biotechnology and chemistry. Yet in many developing countries it is hard to acquire the necessary equipment. Our goal is to re-program a harmless laboratory strain of the E. coli so that the preparation of commonly used proteins, like drugs and enzymes, is controllable by light. To this end we introduce genetic building blocks (of our own design, or ordered from an open-source database)into the bacteria. The project is meant to replace current chemical methods thus making protein production simpler, cheaper and more environmentally friendly. Although everything in nature is renewable and energetically optimized, there are still some obstacles to the efficient use of nature’s own achievements. Up to now, the production and extraction of a protein has required rather expensive equipment and chemicals which can only be provided by a highly specialized high-tech industry. This needs not be so however – nature itself works fine without such help. That is why we wish to develop systems which are in keeping with nature, making its functions available to us – without the additional use of expensive materials if possible. Our approach is based on the great advantage of living systems: self-reproduction. Ideally, most equipment needs for molecular biology could be covered by nature itself. In creating an organism that can be controlled as precisely and as simply as possible in the functions we try to achieve, we hope to gain direct access to nature’s own capabilities to aid us in research.
Who is the Lab in a Cell meant for?
Our main aim is to develop a biological toolkit to be used by scientists for research purposes. Our method enables simple and cheap reproduction of our kit. In placing our results in an open access database, we hope to reach scientists in places without a specialized SynBio industry, helping to improve the working situation on-site. We want our protein expression and purification mechanism to be as independent as possible from high-tech infrastructure. This will facilitate its use in more remote areas of this world for research and diagnostic purposes, enabling scientists to work wherever required with reduced expenditure of resources.
From personal experience we know that in South America, for example, it is sometimes very difficult to obtain the necessary materials, slowing down and complicating the exploration of the Amazonian ecosystems amongst other things.
Our Lab in a Cell concept is not only far simpler to handle than many current technologies, but also cheaper and easier to keep at hand for when needed. We foresee the functions of a synthetic biology lab provided by cells themselves, controlled via what nature provides, such as light for example. Everything needed could fit into a mere suitcase: pipettes and tips, bacterial samples, extracted and dried DNA, a small device for light induction or at least some colour filters. This idea can be extended and modified for basically any cellular function we wish to use for our purposes.