Multiple gene deletion in Cryptococcus neoformans using the Cre-lox system.
Reverse genetics is commonly used to identify and characterize genes involved in a variety of cellular processes. There is a limited set of positive selectable markers available for use in making gene deletions or other genetic manipulations in Cryptococcus neoformans. Here, we describe the adaptation of the Bacteriophage P1 Cre-loxP system for use in C. neoformans, and its application in the excision and reuse of the geneticin drug marker. This tool will allow investigators to make multiple, sequential gene deletions in the same strain, which should facilitate the analysis of multigene families.
School of Science and Technology, Georgia Gwinett College, Lawrenceville, GA, USA. baker@borcim.wustl.edu
Lorina G
LG
Baker
Jennifer K
JK
Lodge
D001287:Attachment Sites, Microbiological / Q000235:genetics*; D017101:Bacteriophage P1 / Q000201:enzymology; D003455:Cryptococcus neoformans / Q000235:genetics*; D017353:Gene Deletion; D005819:Genetic Markers / Q000235:genetics; D005839:Gentamicins / Q000378:metabolism; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed / Q000379:methods*
D005819:Genetic Markers; D005839:Gentamicins; C010680:antibiotic G 418; C045073:Cre recombinase; D019426:Integrases