Presentation on theme: "National Science Foundation, Research Experience for Undergraduates, Utah State University August 5, 2010"— Presentation transcript:
National Science Foundation, Research Experience for Undergraduates, Utah State University August 5, 2010 firstname.lastname@example.org
Problem Pseudomonas Putida KT2440 Key Words Objective Equipment Chemoattractants Growth Curve Phases Results/Data Growth Curve Parameter Velocity Turn Angle X vs. Y Displacement Project Contribution Conclusion Future Work Acknowledgements References Questions
Toxic Chemicals In SoilEffects Due to pesticides, oil spills, landfill leaching etc; soil is contaminated with toxic chemicals: 2,4-Dichlorophenoxyacetic acid Most widely used herbicide in the world Major component in Agent Orange Toluene Naturally occurring in crude oil Byproduct of gasoline production Napthalene Nitrobenzene Water Contamination Farmland Contamination Environmental Damage Health Risks
Chemotaxis: The observable reaction of an organism moving toward or away from the concentration of a certain chemical. Chemoattractant: A certain chemical that the organism of interest is attracted to. Video Microscopy: Microscopic analysis of nano sized samples to capture real time videos: 15 frames/second
Motile due to flagella Non-pathogenic: Safe to be released into the environment Chemotactic towards soil pollutants ◦ Naphthalene ◦ Toluene ◦ Certain aromatic compounds 4-hydroxybenzoate ◦ 2,4-Dichlorophenoxyacetic acid Degrades certain toxic chemicals http://genome.jgi-psf.org/psepu/psepu.home.html
Taken by Serena Bennett with IX71 inverted microscope at 60X magnification at 15 frames/sec.
Observe P. Putida characteristic reactions/indicators from exposure to two separate chemoattractants: Succinic Acid (Succinate) Sodium Benzoate Compare Growth Curve Parameters Velocity (Individual bacterium) T urn Angle (Individual bacterium) Displacement (Individual bacterium)
Spectrophotometer ◦ Measures absorbance (optical density) Proportional to concentration IX71 Inverted Video Microscope ◦ (Olympus, Center Valley, PA, USA) ◦ Digital CCD camera (Olympus DP30BW ImagePro v. 6.1 ◦ (Media Cybernetics, Bethesda, MD, USA) ◦ Image Analysis OriginPro v. 7.5 ◦ (OriginLab Northampton, MA, USA) PPKT2440 ◦ Lysogeny Broth http://www.olympusamerica.com/seg_section/product. asp?product=1023&p=72
Succinic AcidSodium Benzoate 15mM ◦ 88.5mg in 50ml LB ◦ Autoclave 25mM ◦ 180mg in 50ml LB ◦ Autoclave Concentrations taken from Nisar Ahmed Kanhar experiment, Utah State University.
General Phases Phases Compared in KT2440 Project Early Logarithmic Mid Logarithmic Late Logarithmic Early Stationary
OriginPro ◦ Sigmoid Fit ◦ Modified Gompertz Equations 1) 2) 3) 4) OriginPro fitted growth curve of PPKT2440 with no chemoattractant.
OriginPro growth curve of PPKT2440 with 25mM Benzoate OriginPro growth curve of PPKT2440 with 15mM Succinate
A=a-asymptote-highest point in curve k=rate coefficient X c = Center point of inflection µ m =Maximum specific growth rate λ = Lag Phase
15mM Succinate25mM Benzoate Decreased velocity Altered turn angle distribution-bimodal Decrease in overall distance traveled Extended lag phase Lengthens distance traveled in mid log phase Higher average direction change than without chemoattractant Highest maximum absorbance
Find new sensitive indicators More frames/sec in video analysis Reactions to other chemoattractants Research specific pseudomonas genes Mutate genes to create custom bioremediator
Funds from National Science Foundation ◦ Research Experience for Undergraduates Dr. YangQuan Chen ◦ Program Director Dr. Anhong Zhou ◦ Molecular and Cellular Sensing and Imaging Laboratory Dr. Charlie Miller ◦ P. Putida cells Mike Davis ◦ Technical skills
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