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DNA, which contains the genetic information of an organism, consists of long chains of nucleotides. In order to study the functions based on the sequence of these building blocks, DNA molecules must be inserted in carrier molecules (plasmid-vectors) to be multiplied. For this cloning process, a research team from the University of Bayreuth has developed a highly efficient, fast and inexpensive method that is versatile enough to be deployed in all areas of biology, biochemistry and biotechnology. A key feature of the method is that it makes any painstaking screening of bacterial colonies unnecessary. The scientists presented their innovation in the journal Scientific Reports.
One thing that all methods subsumed under the term molecular cloning have in common is that the DNA fragments of interest are first incorporated into larger carrier molecules, the plasmid-vectors. Bacteria are then made to take up these vectors bearing the DNA fragments.
Bacteria - Reproduce - Form - Colonies - DNA
As the bacteria reproduce and form bacterial colonies, the DNA fragments are multiplied thousands of times. Up to now, however, these methods have had one considerable drawback: Since the insertion of DNA fragments into the carrier molecule does not always proceed as smoothly and perfectly as required, only some, but by no means all, colonies come to possess the vectors with the DNA fragments to be duplicated. In order to identify these "success stories," time-consuming and expensive screening has been unavoidable up to now.
The Bayreuth researchers led by Prof. Dr. Stefan Schuster have now succeeded in making this screening redundant. The vector they used is a plasmid that contains a toxic gene. DNA fragments are then incorporated into the plasmid in such a way as to replace this very gene. If this does not succeed, the plasmid keeps its toxic potential. And if, in turn, this plasmid is taken up by an E. coli-bacterium, its...
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