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 Previous Article

Applied and Environmental Microbiology, July 2008, p. 4241-4245, Vol. 74, No. 13
0099-2240/08/$08.00+0     doi:10.1128/AEM.00940-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Highly Efficient Method for Introducing Successive Multiple Scarless Gene Deletions and Markerless Gene Insertions into the Yersinia pestis Chromosome{triangledown}

Wei Sun, Shifeng Wang, and Roy Curtiss III*

Center for Infectious Diseases and Vaccinology, The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, Arizona 85287

Received 24 April 2008/ Accepted 5 May 2008

An efficient two-step recombination method for markerless gene deletion and insertion that can be used for repetitive genetic modification in Yersinia pestis was developed. The method combines {lambda} Red recombination and counterselective screening (sacB gene) and can be used for genetic modification of Y. pestis to construct live attenuated vaccines.

* Corresponding author. Mailing address: The Biodesign Institute, Center for Infectious Diseases and Vaccinology, Arizona State University, P.O. Box 875401, 1001 S. McAllister Avenue, Tempe, AZ 85287-5401. Phone: (480) 727-0445. Fax: (480) 727-0466. E-mail: rcurtiss{at}asu.edu

{triangledown} Published ahead of print on 16 May 2008.

Applied and Environmental Microbiology, July 2008, p. 4241-4245, Vol. 74, No. 13
0099-2240/08/$08.00+0     doi:10.1128/AEM.00940-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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