Can I read Gene disruption by plasmid integration in Listeria monocytogenes: Insertional inactivation of the listeriolysin determinant IisA on EtoBox?

Gene disruption by plasmid integration in Listeria monocytogenes: Insertional inactivation of the listeriolysin determinant IisA by Michael D. Wuenscher; Stephan Köhler; Werner Goebel; Trinad Chakraborty is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Gene disruption by plasmid integration in Listeria monocytogenes: Insertional inactivation of the listeriolysin determinant IisA about?

A plasmid integration technique was developed for insertional inactivation of chromosomal Listeria monocytogenes genes. A Listeria-Escherichia coli shuttle vector (pLSV1) was constructed which carried the temperature-sensitive gram-positive replication origin from plasmid pTV32(Ts). An internal fragment of the listeriolysin gene (lisA) was cloned into pLSV1 to create pLSV2. In L. monocytogenes pLSV2 transformants, plasmid pLSV2 integrated into the L. monocytogenes chromosome at a frequency of 2 x 10(-3) via lisA homology and these cells could be selected at 42 degrees C using a plasmid-encoded erythromycin resistance. Plasmid integration resulted in disruption of the lisA gene, production of a truncated, immunologically cross-reactive listeriolysin protein and loss of the hemolytic phenotype. An improved Listeria protoplast transformation method is also described which facilitates genetic manipulation of Listeria species.

Who reads Gene disruption by plasmid integration in Listeria monocytogenes: Insertional inactivation of the listeriolysin determinant IisA?

Author

Michael D. Wuenscher; Stephan Köhler; Werner Goebel; Trinad Chakraborty

Publisher

Springer; Springer-Verlag; Springer Verlag; Springer Science and Business Media LLC (ISSN 0026-8925)

Published

1991

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)