Dataset Available

Aryl Hydrocarbon Receptor signaling in response to 30 oil sub-fractions

Authors: C. Miller and E. Overton

Published On: Apr 11 2017 19:18 UTC

DOI: https://doi.org/10.7266/N7ZK5F3K

UDI: R5.x280.000:0008

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Project Information

Funded By:
Gulf of Mexico Research Initiative

Funding Cycle:
RFP-V

Research Group:
Toxicological Properties of Specific Aromatic Hydrocarbons Isolated from Fresh and Aged Crude Oil from the Deepwater Horizon Spill

Point of Contact

Charles Miller
Tulane University / School of Public Health and Tropical Medicine
rellim@tulane.edu

Theme keywords

oil, aryl hydrocarbon receptor, bioassay, sub-fraction, toxicity

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Abstract:

This is a lacZ reporter gene bioassay that measures aryl hydrocarbon receptor signaling in response to aromatic hydrocarbons. Here, fractions from the Overton laboratory prepared on 1-20-2017 (13 sub-fractions) and 1-26-2017 (17 sub-fractions) were tested at 3 dilutions for bioactivity. The oil sub-fractions were compared by % normalization to signaling of the positive reference ligand beta-naphthoflavone which was set to 100%. Signaling at greater dilutions indicates greater potency and/or greater chemical content in the sub-fractions.

Suggested Citation:

C. Miller and E. Overton. 2017. Aryl Hydrocarbon Receptor signaling in response to 30 oil sub-fractions. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N7ZK5F3K

Publications:

Fox, J. E., Burow, M. E., McLachlan, J. A., & Miller, C. A. (2008). Detecting ligands and dissecting nuclear receptor-signaling pathways using recombinant strains of the yeast Saccharomyces cerevisiae. Nature Protocols, 3(4), 637–645. doi:10.1038/nprot.2008.33

Purpose:

Identify bioactive sub-fractions of oil.

Data Parameters and Units:

Absorbance at 600 nm (proportional to cell density #cells/mL), Absorbance at 405 nm, Plate number, sample dilution, LacZ units (enzymatic activity, unitless).

Methods:

Quantitative expression of a gene that is regulated by specific oil chemicals. Template of fractionated oil samples included. Samples assayed in triplicate. Samples numbered by date-fraction#. Samples dissolved in 100 uL DMSO. 3 untreated and 3 positive controls included. Methods used are from Fox, J. E., Burow, M. E., Mclachlan, J. A., & Miller, C. A., Iii. (2008). Detecting ligands and dissecting nuclear receptor-signaling pathways using recombinant strains of the yeast saccharomyces cerevisiae. Nature Protocols, 3(4), 637-45. doi:http://dx.doi.org/10.1038/nprot.2008.33

Error Analysis:

The % mean (triplicate assay) and SD of the oil compounds are relative to a positive (100% activity) beta-naphthoflavone ligand control.