Orf306, an esterase coded by the Organophosphate degradation (opd) island of Sphingobium fuliginis ATCC27551 induced gene expression profiling in E. coli MG1655
ABSTRACT: Orf306 induces PNP utilization in E. coli MG1655, the gene expression studies were performed to obtain further details about proteins and pathways that are regulated under the influence of Orf306 Organism: Escherichia coli , Agilent Custom Gene Expression Escherichia Coli MG1655 8x15K ( AMADID: 019439) designed by Genotypic Technology Private Limited
Project description:Transcription profiling of E. coli MG1655, representing the effect of G181D/R258C NusA in comparison to WT NusA Agilent one-color experiment,Organism: Escherichia coli ,Agilent Custom Escherichia coli Gene Expression Microarray 8x60K (AMADID: 48792) designed by Genotypic Technology Private Limited. , Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)
Project description:Transcription profiling of E. coli MG1655, representing the effect of G181D/R258C NusA in comparison to WT NusA Agilent one-color experiment,Organism: Escherichia coli ,Agilent Custom Ecoli Gene Expression Microarray 2x400k designed by Genotypic Technology Private Limited. (AMADID:72220)
Project description:Escherichia coli O157 presents a number of specific problems in terms of food safety and public health. It has been found that E. coli O157 is more resistant to a number of the stresses encountered during food production such as heat, pH and osmotic shock. This greater resistance is thought to contribute to the low infectious dose of E. coli O157 (<100 organisms). Moreover, E. coli O157 is associated with debilitating conditions such as haemorrhagic colitis and haemoytic uraemic syndrome, particularly in children and the elderly. We have been studying the stress responses of E. coli O157:H7 (Sakai) and comparing with a commensal strain of E. coli K-12, MG1655. We found that E. coli O157 (Sakai) is more sensitive to oxidative stress than MG1655. A microarray study of these strains treated with sub-lethal concentrations (0.5mg/ml) of menadione revealed big differences in their responses. In E. coli O157 (Sakai), 540 genes responded significantly to the treatment compared to 121 genes in MG1655. One surprising finding from the microarray data was the observation that many iron-transport genes were up-regulated in E. coli O157 (Sakai) whereas relatively few were induced in MG1655 despite the fact that the bacteria were grown in a medium containing ample iron. We speculated that the induction of iron transport genes in an iron-rich medium might have contributed to the enhanced killing of E. coli O157 (Sakai) through triggering of a Fenton reaction. We speculated that the difference in sensitivity to oxidative stress might be due to differences in the intracellular iron content of E. coli O157 and MG1655. We found that E. coli O157 contains ~50% more iron than MG1655 and believe that during oxidative stress, this iron is released by damaged proteins. The greater levels of free iron in E. coli O157 will trigger a greater Fenton reaction that can damage the ferric uptake regulator (Fur), resulting in unregulated iron transport. In MG1655, the lower iron content results in a smaller Fenton reaction, enabling the cellular protection systems to limit damage and protect Fur. Overnight cultures of E. coli O157 (Sakai) and E. coli K-12 MG1655 were grown in Neidhardt's EZ Rich Defined Medium and diluted 1:100 in 50 ml fresh medium in 125 ml Ehrlenmeyer flasks. The cultures were shaken at 37C until the optical density (OD600) reached 0.4. Each culture was divided into 2 equal parts in identical flasks. One flask contained menadione bisulphite to a final concentration of 0.5 mg/ml; the other flask contained an equivalent volume of distilled water. The flasks were shaken for a further 10 mins and then treated with RNAprotect™ to stabilise the mRNA. The experiment was performed 3 times on different days. Six custom-made microarray slides were used in this study; each slide was hybridised with labelled cDNA made from mRNA taken from untreated and treated E. coli O157 (Sakai) or MG1655.
Project description:Global alterations in trimethoprim (TMP) resistant E. coli are unknown. TMP resistant E. coli have been developed in the laboratory over a period of 48 hours by step-wise increase in TMP concentration and gene expression profiles have been obtained to identify genome wide perturbations in trimethoprim resistance. Biological duplicates of wild-type E. coli MG1655, E. coli resistant to 2 ug/ ml TMP (4xR) and E. coli resistant to 16 ug/ml TMP (32xR) were analyzed.
Project description:The Arabidopsis seeds of Col-0 and pad2.1 were grown for three weeks and were further placed in filter paper for 5 min at 4°C and then transferred to 1/2MS media with 30% PEG at 4°C for an additional 6 hours; the leaves were collected for RNA isolation. Trancriptomic profiling of the combined stress treated Col-0 and pad2.1 indicated downregulation of various abiotic and oxidative stress responsive genes in pad2.1 in response to combined stress treatment. Agilent one-color experiment,Organism: Arabidopsis thaliana ,Agilent Custom Arabidopsis 8x60k Microarray designed by Genotypic Technology Private Limited (AMADID: 48015)
Project description:Investigation of whole genome gene expression level changes in a Escherichia coli MG1655 K-12 ?fnr mutant, compared to the wild-type strain. The mutations engineered into this strain produce a strain lacking the FNR protein. WT strains were grown under aerobic and anaerobic growth conditions. A six chip study using total RNA recovered from two separate cultures of Escherichia coli MG1655 K-12 WT (aerobic and anaerobic) and two separate cultures of the ?fnr mutant strain (anaerobic). Each chip measures the expression level of 4,661 genes from Escherichia coli MG1655 K-12 with eight 60-mer probes per gene, with each probe represented twice on the array.
Project description:Investigation of whole genome gene expression level changes in a Escherichia coli MG1655 K-12 ∆hns/∆stpA strain from exponental growth under aerobic and anaerobic growth conditions. The results are further described in the article Genome-scale Analysis of E.coli FNR Reveals the Complex Features of Transcrtipion Factor Binding. A four chip study using total RNA recovered from two separate cultures of Escherichia coli MG1655 K-12 ∆hns/∆stpA mutant strain under aerobic and anaerobic growth conditions. Each chip measures the expression level of 4,661 genes from Escherichia coli MG1655 K-12 using a high-density tiling array consisting of ~385,000 60mer probes spaced every 12 bp.
Project description:Microarray analysis of the gill tissues of WSSV infected shrimp (P. monodon) at different time intervals 6 hrs, 24 hrs, 48 hrs and moribund stage of post WSSV infection was carried out to identify differentially expressed genes in response to WSSV infection. The shrimps in WSSV challenege experiment were challenged through intra muscular route with known concentration of virus. The important immune genes identified would be further characterized by sequence analysis and gene expression profile would be validated by real time PCR One-color experiment,Organism: Penaeus monodon, Custom Penaeus monodon (Black Tiger Shrimp) 8x60k designed by Genotypic Technology Private Limited (AMADID: 041733), Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)