|
| Status |
Public on Aug 15, 2013 |
| Title |
Cryptococcus neoformans H99 extracted from J774A.1 macrophage-like cells |
| Sample type |
SAGE |
| Anchor |
NlaIII
|
| Tag Count |
38544 |
| Tag Length |
17 |
| |
|
| Source name |
Cryptococcus neoformans H99
|
| Organism |
Cryptococcus neoformans var. grubii H99 |
| Characteristics |
mating type: alpha
isolate: human isolate
pathogen: human pathogenic basidomycete yeast
incubation partner: J774A.1 murine macrophage-like cell line
|
| Biomaterial provider |
John Perfect, Duke University, North Carolina, USA
|
| Treatment protocol |
untreated
|
| Growth protocol |
Murine macrophage-like cell line J774A.1 was grown at 37°C in 10% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated fetal calf serum (FBS), 1% nonessential amino acids, penicillin-streptomycin, and L-glutamine (Invitrogen). Cryptococcus cells were opsonized with monoclonal antibody 18B7 against capsule and macrophages were treated with recombinant mouse gamma interferon (IFN-gamma) and lipopolysaccharide (LPS) prior to co-incubation at a multiplicity of infection (MOI) of 1:1. Macrophages were inoculated with H99 cells and washed after 1 h of inoculation to remove unattached, extracellular fungal cells. After 6 h of incubation, sterile, ice-cold distilled H2O was applied to each well to lyse the macrophages, and the fungal cells (4 x 107) were harvested by centrifugation.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
polyA RNA was extracted using standard TRIZOL procedures and libraries were constructed using the NlaIII anchoring enzyme and the I-Long SAGE kit (Invitrogen).
|
| |
|
| Description |
Cryptococcus neoformans H99
|
| Data processing |
Tag detection and counting was carried out using PHRED and CrossMatch software (University of Oklahoma’s Advanced Center for Genome Technology, http://www.genome.ou.edu/cneo.html)
|
| |
|
| Submission date |
Aug 15, 2012 |
| Last update date |
Aug 15, 2013 |
| Contact name |
Emma Griffiths |
| E-mail(s) |
griffie@msl.ubc.ca
|
| Organization name |
University of British Columbia
|
| Department |
Michael Smith Laboratories
|
| Lab |
Kronstad
|
| Street address |
2185 East Mall
|
| City |
Vancouver |
| State/province |
BC |
| ZIP/Postal code |
V6T 1Z4 |
| Country |
Canada |
| |
|
| Platform ID |
GPL15941 |
| Series (1) |
| GSE41066 |
A defect in ATP-citrate lyase links acetyl-CoA production, virulence factor elaboration and virulence in Cryptococcus neoformans |
|
