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Comparative expression analysis between logarithmic and stationary phase of T. thermophilus HB8 on a synthetic medium

ABSTRACT: We observed the expression profile of the total mRNA of Thermus thermophilus HB8 wild-type strain grown in a synthetic medium at 70°C. Three wild-type T. thermophilus HB8 strains were pre-cultured at 70°C for 16 h in 3 ml of synthetic medium. The cells (2 ml) were inoculated into 500 ml of the same medium and then cultivated at 70°C. Cells were collected after 10 h and 20 h, and then crude RNA was extracted. The expression of each mRNA at each time point was analyzed on a GeneChip as described under Sample Description Sheet of each sample in this website.

ORGANISM(S): Thermus thermophilus HB8

SUBMITTER: Yoshihiro Agari

PROVIDER: E-GEOD-11671 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Global gene expression mediated by Thermus thermophilus SdrP, a CRP/FNR family transcriptional regulator.

Agari Yoshihiro Y Kashihara Aiko A Yokoyama Shigeyuki S Kuramitsu Seiki S Shinkai Akeo A

Molecular microbiology 20080811 1

Thermus thermophilus SdrP is one of four cyclic AMP receptor protein (CRP)/fumarate and nitrate reduction regulator (FNR) family proteins from the extremely thermophilic bacterium T. thermophilus HB8. Expression of sdrP mRNA increased in the stationary phase during cultivation at 70 degrees C. Although the sdrP gene was non-essential, an sdrP-deficient strain showed growth defects, particularly when grown in a synthetic medium, and increased sensitivity to disulphide stress. The expression of se ...[more]

PMID: 18699868

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Project description:We compared the expression profile of TTHA1939 deletion mutant strain of Thermus thermophils HB8 with that of wild-type. The mutant strain grown in minimum essential medium (CS medium) exhibited growth arrest and cellular aggregation during the logarithm growth phase. In this phase, 99 genes were suppressed and 63 genes were activated for the mutant strain. The suppressed genes included genes of ribosomal proteins, TCA cycle, amino acid biosynthesis, fatty acid biosynthesis, cobalamin biosynthesis, transporters, and cell division including dnaB and ftsZ. The activated genes included genes of nitrogen metabolism, stress proteins, and proteases. We suggested that these transcriptional alterations in the mutant cell were induced by the ppGpp accumulation which was prevented by ppGpp degradation activity of TTHA1939 in WT cell. Keywords: gene deletion, minimum essential medium, logarithm growth phase Wild-type and the mutant strains were precultured in rich medium (TR medium) for two times at 70 oC and then subcultured to minimum essential medium (CS medium). These cells were harvested during the logarithm growth phase (800 minutes from the start of culture). Total RNA were extracted from each strain and used for the cDNA synthesis. For the biological replication, above experiments were performed four times independently. The cDNA fragments were labeled with biotin-dideoxy UTP, using terminal transferase according to the manufacturerâs instructions (ENZO Biochem. Inc., Farmingdale, NY). The 3â-terminal-labeled cDNA (2 micro g) was hybridized to a TTHB8401 GeneChip (Affymetrix, Santa Clara, CA). The Probe Array was scanned with a GeneArray Scanner (Affymetrix), and then, the image data was scaled to the target intensity by one-step Tukeyâs biweight algorithm using GeneChip Operating software, version 1.0 (Affymetrix, Santa Clara, CA). The data analysis was performed by using GeneSpring GX (Agilent Tech.). The genes which had detection call of âpresenceâ more than 4 times from 8 samples were used for following analysis. The normalized intensities for the mutant strains were calculated by using the intensities for wild-type strains of each data set, and then, the average of these intensities were used. The genes which produced P value < 0.1 in the Studentâs t-test were extracted. Among them, the genes whose expression level was different more than 2 fold between two strains were considered as significant.

Project description:We compared the expression profile of TTHA0118 deletion mutant strain of Thermus thermophils HB8 with that of wild-type. The mutant strain grown in minimum essential medium (CS medium) exhibited growth retardation. In early log phase, 29 genes were suppressed and 25 genes were activated for the mutant strain. The suppressed genes included genes of electron transport, and cell growth and division. The activated genes included genes of stress response. We suggested that these transcriptional alterations in the mutant cell were induced by the pAp accumulation and mononucleotides starvation. Keywords: cell type comparison Keywords: cell type comparison Wild-type and the mutant strains were precultured in rich medium (TT medium) for two times at 70 oC and then subcultured to minimum essential medium (CS medium). These cells were harvested at 7 × 10^8 cells/ml. Total RNA were extracted from each strain and used for the cDNA synthesis. For the biological replication, above experiments were performed four times independently. The cDNA fragments were labeled with GeneChip DNA Labeling Reagent (Affymetrix, Santa Clara, CA), using terminal transferase according to the manufacturer's instructions. The 3'-terminal-labeled cDNA (2 micro g) was hybridized to a TTHB8401 GeneChip (Affymetrix). The Probe Array was scanned with a GeneArray Scanner (Affymetrix), and then, the image data was scaled to the target intensity by one-step Tukey's biweight algorithm using GeneChip Operating software, version 1.0 (Affymetrix). The data analysis was performed by using GeneSpring GX (Agilent Tech.). The genes which had detection call of 'presence' more than 4 times from 8 samples were used for following analysis. The normalized intensities for the mutant strains were calculated by using the intensities for wild-type strains of each data set, and then, the average of these intensities were used. The genes which produced P value < 0.05 in the Student's t-test were extracted. Among them, the genes whose expression level was different more than 2 fold between two strains were considered as significant.

Dataset Information

Comparative expression analysis between logarithmic and stationary phase of T. thermophilus HB8 on a synthetic medium

Publications

Global gene expression mediated by Thermus thermophilus SdrP, a CRP/FNR family transcriptional regulator.

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