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Applied and Environmental Microbiology, September 2001, p. 4361-4364, Vol. 67, No. 9
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.9.4361-4364.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Efficient System for Directed Integration into the Lactobacillus acidophilus and Lactobacillus gasseri Chromosomes via Homologous Recombination

W. M. Russell1 and T. R. Klaenhammer1,2,*

Department of Microbiology1 and Department of Food Science,2 Southeast Dairy Foods Research Center, North Carolina State University, Raleigh, North Carolina 27695

Received 22 February 2001/Accepted 30 May 2001

An efficient method is described for the generation of site-specific chromosomal integrations in Lactobacillus acidophilus and Lactobacillus gasseri. The strategy is an adaptation of the lactococcal pORI system (K. Leenhouts, G. Venema, and J. Kok, Methods Cell Sci. 20:35-50, 1998) and relies on the simultaneous use of two plasmids. The functionality of the integration strategy was demonstated by the insertional inactivation of the Lactobacillus acidophilus NCFM lacL gene encoding beta -galactosidase and of the Lactobacillus gasseri ADH gusA gene encoding beta -glucuronidase.

* Corresponding author. Mailing address: North Carolina State University, Department of Food Science, Box 7624, Raleigh, NC 27695. Phone: (919) 515-2972. Fax: (919) 515-7124. E-mail: klaenhammer{at}ncsu.edu.

Applied and Environmental Microbiology, September 2001, p. 4361-4364, Vol. 67, No. 9
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.9.4361-4364.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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