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What makes a chromosome a chromosome? This is the central question to be answered by the research we do in our group. Chromosomes need to be copied before division of the cell (replication), they must be distributed to the daughter cells (segregation) and they need to be folded since they are much longer than the cell itself. Without systems that realize these fundamental processes no chromosome would work. We study the molecular details of such systems using the bacterial model organisms Escherichia coli and Vibrio cholerae. E. coli is so useful because it is the best studied bacterium. V. cholerae, the causative agent of the cholera disease, is particularly interesting because it has two chromosomes unlike most other bacteria. By studying these relatively simple model organisms we hope to understand fundamental mechanisms underlying chromosome function in all living organisms.
Systems for chromosome maintenance usually consist of a DNA binding protein and the respective DNA sequence motive. The distribution of such DNA motives on the chromosome often determines the function of the system.
The possibility to study such systems using designed motive distributions on synthetic chromosomes bears an enormous potential. Our plan is to establish secondary synthetic chromosomes beside the main chromosome in E. coli. The biological template is V. cholerae which has two natural chromosomes. The synthetic chromosomes should have a size of about 100.000 base pairs. This is a multiple of the cell size and allows analysis of chromosome folding which would not be necessary for small DNA molecules.
Only recently an American research group for the first time synthesized a complete bacterial chromosome. The technical challenge is enormous. First, smaller DNA pieces are synthesized and then assembled stepwise to build whole chromosomes. For this work we are planning to use the lab automation platform of the Becker group.
The synthetic chromosomes will be used to understand basic mechanisms that make chromosomes function. In the future this basic research will help to design and construct tailored chromosomes for diverse applications.
We are entering an era in life science where synthesis of chromosome size molecules becomes feasible. Do we actually understand construction rules of natural chromosomes completely? This would be essential to design and build synthetic chromosomes for organisms with new capabilities. We study bacterial chromosome biology using a variety of methods with emphasis on synthetic biology approaches.
1. You have to understand chromosome biology to build synthetic chromosomes.
2. You have to build synthetic chromosomes to understand chromosome biology.
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