|
| Status |
Public on Jun 11, 2009 |
| Title |
JB1ts_x_JB2ts_5dpi_9h_31C_III |
| Sample type |
RNA |
| |
|
| Source name |
Tumor material from maize leaves 5 days post RAb1ts/RAb2ts mixed infection at 31°C
|
| Organism |
Mycosarcoma maydis |
| Characteristics |
strain: RAb1ts (a1,bW1bE1ts) and RAb2ts (a2,bW2bE2ts)
|
| Biomaterial provider |
Ramon Wahl and Jörg Kämper
|
| Treatment protocol |
U. maydis RAb1ts/RAb2ts infected maize plants (Early Golden Bantam) were grown in a phytochamber in a 15h/9h light-dark cycle; light period started/ended with 1h ramping of light intensity. Prior to infection with U. maydis maize plants were kept at 28 °C (light) and 20 °(dark). Plantlets were individually sown in pots with potting soil (Fruhstorfer Pikiererde) and infected 7 days after sowing, 1 h before end of the light period. After mixed infection with RAb1ts/RAb2ts the plants were kept at 22 °C. 111 hours post infection plants were shifted for 9 hours to 31 °C, inactivating b function after RAb1ts/RAb2ts but not after FB1/FB2 infection. Infected leaf tumor material from at least 10 plants was collected 5 days post infection, 1 h before the end of the light period. Sample material from at least 3 independent replicates of RAb1ts/RAb2ts infections at 31°C was directly frozen in liquid nitrogen for RNA-extraction.
|
| Growth protocol |
U. maydis strains were grown at 28°C in YEPS (Tsukuda et al., 1988) medium as pre-culture for plant infections. Plant infections were performed as described by (Brachmann et al., 2001).
- Brachmann, A., Weinzierl, G., Kämper, J., and Kahmann, R. (2001) Mol Microbiol 42(4): 1047-63.
- Tsukuda, T., Carelton, S., Fotheringham, S., and Holloman, W.K. (1988). Mol Cell Biol 8: 3703-09.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the TRIZOL-reagent (Invitrogen) according to the manufacturer's instructions. RNA samples were further column purified using (Qiagen RNeasy) and the quality checked using a Bioanalyzer with a RNA 6000 Nano LabChip kit (Agilent).
|
| Label |
biotin/phycoerythrin
|
| Label protocol |
Target preparation was performed according to the standard Affymetrix protocol using the Enzo BioArray RNA transcript labeling kit, except for a reaction temperature of 50 °C for first strand cDNA synthesis
|
| |
|
| Hybridization protocol |
Affymetrix EukGE-2V4 on GeneChip Fluidics Station 400
|
| Scan protocol |
Affymetrix GSC3000
|
| Description |
Experiment to measure effects after temperature-mediated (31 °C) inactivation of the b heterodimer in planta in RAb1ts/RAb2ts. Control Experiment is FB1/FB2 wildtype infection under restrictive conditions. RAb1ts (a1,bW1bE1ts) and RAb2ts (a2,bW2bE2ts) are compatible strains that carry temperature sensitive bE allels, which is active under permissive conditions (22°C) and inactive under restrictive conditions (31°C). The mating types are indicated in brackets. The strains are derivatives of JB1 (a1,∆b) and JB2 (a2,∆b) with integration of bW/bEts into the ip-locus (carboxin resistance).
|
| Data processing |
MAS5.1
|
| |
|
| Submission date |
Jun 08, 2009 |
| Last update date |
Jun 10, 2009 |
| Contact name |
Jörg Thomas Kämper |
| E-mail(s) |
joerg.kaemper@kit.edu
|
| Phone |
+49 721 608 5670
|
| Organization name |
Karlsruhe Institute of Technology
|
| Department |
Department for Applied Biosciences
|
| Street address |
Hertzstrasse 16, Geb. 06.40
|
| City |
Karlsruhe |
| ZIP/Postal code |
76187 |
| Country |
Germany |
| |
|
| Platform ID |
GPL3681 |
| Series (1) |
| GSE16501 |
Temperature-mediated inactivation of the b heterodimer during biotrophic development of U. maydis |
|
