| Description: | CK2 is an essential protein kinase involved in a wide variety of cellular processes. The aim of these experiments was to decipher the role of CK2 kinase in Arabidopsis DNA damage response. It is well reported that a rapid transcriptional burst occurs when plants are exposed to gamma radiation. For this reason, we studied the global changes of gene expression in 7-day-old Arabidopsis seedlings expressing a dominant-negative mutant of CK2 (CK2mut plants). Those plants contain a CK2-alfa inactive subunit cloned downstream of a GVG inducible promoter, which is inducible by Dexametahone (Dex)(Moreno-Romero et al. 2008). The gene expression changes were analyzed in gamma-irradiated, Dex-treated mutant seedlings, and were compared to those in irradiated, Dex-untreated seedlings.METHOD: Arabidopsis seeds were surface sterilized, cold-treated at 4ºC for 48 hours, and then sown on MS plates and grown for five days. Afterwards, plantlets were transferred to MS plates supplemented with 1uM Dex and after two days were treated with 100 Gy of gamma rays. Total RNA was isolated from frozen plantlets with Trizol (Invitrogen) and later purified using Qiagen RNA easy Plant Mini Kit columns (Qiagen, Hilden, Germany). RNA quality was checked using Experion (Bio-Rad laboratories). Three independent RNA preparations were made from CK2mut pooled samples in two different conditions: Dex-treated and irradiated, or mock-treated and irradiated. The data relative to controls necessary for this experiment (plants transformed with the empty vector, and non-irradiated plants) are deposited in NASC (ref. NASCARRAYS-642).Moreno-Romero et al. 2008. A role for protein kinase CK2 in plant development: evidence obtained using a dominant-negative mutant. The Plant Journal 55, 118-130 [2008]. |
NASCArrays Information at The BAR
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