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Applied and Environmental Microbiology, July 2005, p. 4097-4100, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.4097-4100.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Multiple-Mutation Reaction: a Method for Simultaneous Introduction of Multiple Mutations into the glpK Gene of Mycoplasma pneumoniae

Claudine Hames, Sven Halbedel, Oliver Schilling, and Jörg Stülke*

Department of General Microbiology, Institute of Microbiology and Genetics, Georg-August University Göttingen, Grisebachstr. 8, D-37077 Göttingen, Germany

Received 19 November 2004/ Accepted 4 January 2005

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.

* Corresponding author. Mailing address: Department of General Microbiology, Institute of Microbiology and Genetics, Georg-August University Göttingen, Grisebachstr. 8, D-37077 Göttingen, Germany. Phone: 49-551-393781. Fax: 49-551-393808. E-mail: jstuelk{at}gwdg.de.

Applied and Environmental Microbiology, July 2005, p. 4097-4100, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.4097-4100.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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