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Series GSE10974 Query DataSets for GSE10974
Status Public on Apr 14, 2008
Title Expression profiles of E. coli groESL(-) cells expressing human Hsp60(wt)/Hsp10 or Hsp60-(p.Val98Ile)/Hsp10 operons
Platform organism Escherichia coli K-12
Sample organism Escherichia coli
Experiment type Expression profiling by array
Summary To identify regulatory responses that could give clues as to the cause(s) of the growth arrest observed in E. coli cells expressing mutant Hsp60-(p.Val98Ile) and Hsp10 versus cells expressing wild type Hsp60 and Hsp10, we performed microarray analyses. We reasoned that the decreased chaperonin activity of the Hsp60-(p.Val98Ile) protein would greatly impair or abolish folding of E. coli proteins that depend on chaperonin assistance. The resulting decreased levels of various activities may cause compensatory up-regulation or deregulation of genes. The transcriptomes of cells shifted to expression of the arabinose-inducible chaperonin operon with either the Hsp60-(p.Val98Ile) mutant or the wild type Hsp60 from the second plasmid were compared. RNA samples from three independent cultures expressing the mutant or two independent cultures expressing wild type Hsp60 were pooled, respectively.
Keywords: Comparison of cells expressing two different variants of Hsp60
 
Overall design B178 (groELS)-deleted cells double-transformed with two plasmids harboring an IPTG-inducible operon with wild type human Hsp60 and Hsp10 and a second plasmid harboring an arabinose inducible operon with either Hsp60-(p.Val98Ile) and Hsp10 (mutant cells) or wild type Hsp60 and Hsp10 (wild type cells) were grown in dYT medium (21) supplemented with kanamycin (10 mg/L), ampicillin (100 mg/L) and IPTG (0.1mM) at 30°C in a shaking water bath. At OD536 ≈ 0.1 bacteria were harvested by centrifugation at ambient temperature, resuspended in fresh medium, and 0.2% arabinose was added. Growth was continued at 30°C. After 10 hours 3ml samples were withdrawn and RNA was isolated.
Two independent transformant clones of the wild type cells and three of the mutant cells were grown in this way. The RNA preps from the 2 wild type and the three mutant cells, respectively, were pooled, cDNA was synthesized, labelled and subjected to Affymetrix arrays.
 
Contributor(s) Bross P, Naundrup S, Hansen J, Nielsen MN, Christensen JH, Kruhøffer M, Palmfeldt J, Corydon TJ, Ang D, Georgopoulos C, Gregersen N, Nielsen KL
Citation(s) 18400758
Submission date Mar 28, 2008
Last update date Mar 19, 2012
Contact name Peter Bross
E-mail(s) peter.bross@ki.au.dk
Phone +4589495147
Fax +4589496018
Organization name Aarhus University Hospital, Skejby
Department Institute for Clinical Medicine
Lab Research Unit for Molecular Medicine
Street address Brendstrupgaardsvej 100
City Århus
ZIP/Postal code 8200
Country Denmark
 
Platforms (1)
GPL199 [Ecoli_ASv2] Affymetrix E. coli Antisense Genome Array
Samples (2)
GSM277818 E.coli/Hsp60-wt/wt(-/+)
GSM277819 E.coli/Hsp60-wt/VAL98Ile(-/+)
Relations
BioProject PRJNA107175

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Supplementary file Size Download File type/resource
GSE10974_RAW.tar 2.8 Mb (http)(custom) TAR (of CEL, CHP)
SRA Run SelectorHelp
Raw data provided as supplementary file
Processed data included within Sample table

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