Team:WLC-Milwaukee/Background

PLACEHOLDER

WLC iGEM 2018 | Background

BACKGROUND

Introduction

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Water Contamination and Testing (United States and Global)

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Lambda Phage

Lambda phage is a bacteriophage that can infect the well-known bacteria Escherichia coli. It was discovered in 1950 by microbiologist, Esther Lederberg, whose finding would lead to further investigation on gene regulation and recombination [1]. Lambda phage is composed of an icosahedral head and a flexible tail. Lambda phage infects E. coli by using its J protein on its tail to recognize the receptor integral outer membrane protein, LamB, which is found on the surface of E.coli. Research conducted by Karine Gibbs showed that LamB is not distributed uniformly throughout the outer membrane of E.coli, and in fact its distribution changes readily and quickly, illustrating the fluidity of the cell membrane [2]. Upon binding to the surface of E.coli, Lambda phage is able to inject its 48,502 bp of DNA into E. coli where it can either replicate or become a prophage [3]. This means that Lambda phage is capable of following the lytic pathway if it replicates, or the lysogenic pathway if it inserts its DNA into E. coli’s chromosome.


Lambda Phage

The discovery of Lambda phage has many important applications which have been utilized in past years and also continues to be researched on. For example, the water testing kit we are composing relies heavily on utilizing the binding specificity of lambda phage to E.coli in order to rapidly detect E. coli in water samples. Another use of Lambda phage is to use it for phage therapy, which simply means that Lambda phage is used to attack and kill E.coli [4]. More broadly, lambda phage is used as a model to investigate the mechanisms of biological processes, such as the specific binding of bacteriophage to bacteria. The importance of Lambda phage and its discovery has implications in both the health sciences and the environment, making it one of the most important bacteriophages in biotechnology.

  • [1] http://www.whatisbiotechnology.org/index.php/people/summary/Lederberg_Esther
  • [2] Gibbs, K. A., Isaac, D. D., Xu, J. , Hendrix, R. W., Silhavy, T. J. and Theriot, J. A. (2004), Complex spatial distribution and dynamics of an abundant Escherichia coli outer membrane protein, LamB. Molecular Microbiology, 53: 1771-1783. doi:10.1111/j.1365-2958.2004.04242.x
  • [3] https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3509688/
  • [4] https://brainmass.com/biology/cell-and-molecular-biology/lambda-phage-biotechnology-575620

Horse Radish Peroxidase

Horseradish peroxidase (HRP) is an enzyme used in molecular biology. It can be used in various biological methods, such as ELISA, Western Blotting, Southern Blotting, and EMSA [1]. HRP’s colorimetric and chemiluminescent properties make it a useful agent in many tests. HRP acts as a catalyst in redox reactions [3]. A luminol and H2O2 solution can release light for a very long time, but when HRP is added, it usually reaches it maximum light emission between 1 and 5 minutes [4]. This makes it a useful detection method. HRP can be used in more qualitative methods as well, since it induces changes in color when used with chromogenic assays. Commonly used substrates for this type of detection are 3,3',5,5'-tetramethylbenzidine (TMB), 2,2' -azino-di-[3-ethylbenzthiazoline-6-sulfonic acid] (ABTS), and diaminobenzidine (DAB) [3,4]. HRP when added to TMB, for example, yields a 2-electron oxidation product which yields a blue color. HRP is also up and coming in the medical field. When combined with other agents, it is heavily reactive toward tumor cells in humans [2]. It also serves as a functional tracer. HRP can be injected into the bloodstream and then identified at different places in the pathway using the DAB/H2O2 reaction [4]. This makes HRP very useful for immunocytochemistry and electron microscopy [4].


Horse Radish Peroxidase Enzyme Structure

  • [1] https://www.sigmaaldrich.com/life-science/biochemicals/biochemical-products.html?TablePage=15552120
  • [2] https://chem.libretexts.org/Textbook_Maps/Biological_Chemistry/Catalysts/Case_Studies/Horseradish_Peroxidase
  • [3] http://www.bio-rad.com/featured/en/hrp-substrate.html
  • [4] https://www.sciencedirect.com/topics/neuroscience/horseradish-peroxidase
  • [5] http://www.jbc.org/content/257/7/3669.full.pdf

WLC-Milwaukee