Cryptococcus neoformans Genome ORF Knockout Collection Version 1.0 (CnKOv1,0). The Madhani laboratory at the University of California-San Francisco recently generated deletions in approximately 1,200 Open Reading Frames (ORFs) (i.e., about 20% of the genome ORFs) of strain H99 (serotype A, var. grubii) of this organism. Each ORF deletion is made through homologous recombination in which a genome ORF is replaced by a PCR-generated DNA fragment containing selection marker natR and a unique barcode of 40 base pairs. The genotype of each of these gene knockout strains is MATalpha cnx####::natR, wherein x refers to an alphabet and #### to a 4-digit number and natR represents the gene that confers resistance to nourseothricin. For further information on the methodologies in making gene knockouts via homologous recombination and generation of 40-bp unique barcodes, see the publications: Davidson et al., Fungal Genetics & Biology 29: 38-48 (2000); and Nelson et al., Genetics 157: 935-947 (2001).
Strains of Cryptococcus neoformans Genome ORF Knockout Collection (CnKOv1.0) are distributed as a set of fourteen 96-well standard microtiter plates (called CNKO-PS) or single strains can be ordered individually. Please contact us at (800) 638-6597 or firstname.lastname@example.org if you have any questions.