BioProject

We analyzed the expression profile of wild-type strain and ttha1564 deletion mutant strain of Thermus thermophils HB8 under alkylating condition. More...

We analyzed the expression profile of wild-type strain and ttha1564 deletion mutant strain of Thermus thermophils HB8 under alkylating condition. In wild type strain, 107 genes and 96 genes were over 2-fold up- and under 0.5-fold down-regulated by alkylation, respectively. In ttha1564 deletion mutant, 35 genes and 12 genes were over 2-fold up- and under 0.5-fold down-regulated by alkylation, respectively. Keywords: stress response comparison The T. thermophilus HB8 wild-type and delta ttha1564 strains were cultured in TT medium (0.4%(w/v) polypeptone, 0.2%(w/v) yeast extract, 0.1%(w/v) NaCl, 0.4 mM CaCl2, and 0.4 mM MgCl2, (pH 7.5)) at 70°C until the OD600 reached 0.8. The cultures were then divided into two flasks and an equal volume of TT medium prewarmed at 70°C was added. One of the media culture of each strain contained 100 micro g/ml MNNG. We confirmed that the cells can continue to grow in TT medium containing 100 micro g/ml MNNG but DNAs are alkylated sufficiently (Onodera, 2011). After further cultivation at 70°C for 10 min, the diluted cultures were collected into an equal of 100% ethanol and stored at -80°C. The procedures used for extraction of crude RNA, synthesis of cDNA, labeling of cDNA fragment, hybridization to a TTHB8401a520105F GeneChip (Affymetrix) and data analysis were basically the same as those described previously (Agari et al., 2008; Shinkai et al., 2007). To determine the mRNA expression level of each sample, image data for the three independent samples were processed. Overall design: stress response comparison Less...