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Researchers develop new method for facile identification of proteins in bacterial cells

is based on," explained Dr. Halden. "Instead of spending a lot of time and resources on eliminating the background noise to find the signal, our method increases the signal upfront so that it stands out above the background noise. By forcing an up-regulation of enzyme expression in the bacterium of interest, our target can be identified amidst all of the other cell components," he said.

Halden and his colleagues tested their technique using Sphingomonas wittichii strain RW1, the only bacterium known to consume the backbone of toxic polychlorinated dibenzo-p-dioxins and dibenzofurans as a food source. The researchers already knew that feeding dioxins to RW1 would cause an increased enzyme level as the bacterium consumed the model pollutant. Their study shows that this increase can be easily identified by PMF using mass spectrometry.

"Our procedure simplifies the entire identification process," said David Colquhoun, MS, a doctoral fellow with the Johns Hopkins Center for a Livable Future, "With the new tool, we can conveniently and rapidly identify both pollutant-degrading bacteria and their characteristic proteins that effect pollutant transformation."

"This method represents a new investigative tool in bioremediation, which is the science of using biological organisms as a means of decontaminating polluted soils and water," said Dr. Halden.

Johns Hopkins University is seeking partners who would like to license this patent-pending methodology. Inquiries may be directed to Deborah Alper at the Johns Hopkins Bloomberg School of Public Health at dalper@jhsph.edu or 443-287-0402.

"Identification and Phenotypic Characterization of Sphingomonas wittichii Strain RW1 by Peptide Mass Fingerprinting Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry" was written by Rolf U. Halden, David R. Colquhoun and E.S. Wisniewski.


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Source:Johns Hopkins University Bloomberg School of Public Health


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