Multiple-mutation reaction: a method for simultaneous introduction of multiple mutations into the glpK gene of Mycoplasma pneumoniae |
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Authors: | Hames Claudine Halbedel Sven Schilling Oliver Stülke Jörg |
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Affiliation: | Department of General Microbiology, Institute of Microbiology and Genetics, Georg-August University G?ttingen, Grisebachstr. 8, D-37077 G?ttingen, Germany. |
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Abstract: | In Mycoplasma pneumoniae, the UGA opal codon specifies tryptophan rather than a translation stop site. This often makes it difficult to express Mycoplasma proteins in E. coli isolates. In this work, we developed a strategy for the one-step introduction of several mutations. This method, the multiple-mutation reaction, is used to simultaneously replace nine opal codons in the M. pneumoniae glpK gene. |
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