Team:UCLA/Project

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Revision as of 19:45, 26 September 2013

Overall project

In-Vitro Immune System

Diversity is an important feature of biological systems, and it can be harnessed to serve as a useful tool in synthetic biology.

Many organisms have molecular mechanisms to generate diversity at the DNA level in order to accelerate evolution. This is true in particular with host/pathogen interactions, where the host must mutate to protect itself from the pathogen and the pathogen mutates to evade host defenses. One important example of such diversity generating mechanisms is found in the Bordetella bacteriophage BPP-1. Bordetella is a genus of bacteria that can infect the mammalian respiratory tract, causing diseases such as pertussis (whooping cough). The infectious cycle of the Bordetella bacteria involves a switch between a non-infectious and an infectious stage with different virulence factors and surface receptors expressed. Interactions between bacteria surface receptors and phage tail fiber proteins are extremely specific. The tip of each virus tail-fiber is a protein called the major tropism-determinant protein (mtd), which binds to the hosts’ surface and allows the phage to infect it. In order to successfully infect both phases of the host bacteria, the BPP-1 phage uses a complex genetic mechanism to generate diversity at the host-recognition site of the mtd protein.

Though the general structure of the mtd protein is constant and very stable, its active ends are extremely variable. Each phage produces a slightly different structural variant of the protein, with the hope that at least one phage can successfully bind to the host and infect it- akin to the way our immune system produces numerous variants of antibodies so that at least one can bind to the antigen of interest.

[you should put some background info here about how people have done phage display with this system in the past] Our team hopes to develop a fully in-vitro system to express and select for mtd variants of interest.

We plan on first generating a diverse DNA library of the Mtd gene, then using mRNA-display to detect and screen for the variants of the protein that can bind to a selected target. In our case, the target will be the surface of E. Coli bacterial cells.

Retrieved from "http://2013.igem.org/Team:UCLA/Project"

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 <h4>In-Vitro Immune System</h4>
-Diversity is an intrinsic characteristic of nature, and it can be harnessed to serve as a useful tool in synthetic biology.
+Diversity is an important feature of biological systems, and it can be harnessed to serve as a useful tool in synthetic biology.
-In particular, this diversity is exemplified in a species of bacteriophage known as the <i>Bordetella</i> BPP-1 phage. The <i>Bordetella</i> bacteria, a genus of small Gram-negative bacteria, frequently changes its surface topography over the course of its life cycle. To successfully infect its constantly-changing host, the BPP-1 phage uses diversity as its weapon of choice. The tip of each virus tail-fiber is a protein called the major tropism-determinant protein (mtd), which binds to the hosts’ surface and allows the phage to infect it.
+Many organisms have molecular mechanisms to generate diversity at the DNA level in order to accelerate evolution. This is true in particular with host/pathogen interactions, where the host must mutate to protect itself from the pathogen and the pathogen mutates to evade host defenses. One important example of such diversity generating mechanisms is found in the <i>Bordetella</i> bacteriophage BPP-1. <i>Bordetella</i> is a genus of bacteria that can infect the mammalian respiratory tract, causing diseases such as pertussis (whooping cough). The infectious cycle of the <i>Bordetella</i> bacteria involves a switch between a non-infectious and an infectious stage with different virulence factors and surface receptors expressed. Interactions between bacteria surface receptors and phage tail fiber proteins are extremely specific. The tip of each virus tail-fiber is a protein called the major tropism-determinant protein (mtd), which binds to the hosts’ surface and allows the phage to infect it. In order to successfully infect both phases of the host bacteria, the BPP-1 phage uses a complex genetic mechanism to generate diversity at the host-recognition site of the mtd protein.
-Though the general structure of the mtd protein is constant and is very stable, its active ends are extremely variable. Each phage produces a slightly different structural variant of the protein, with the hope that at least one phage can successfully bind to the host and infect it- akin to the way our immune system produces numerous variants of antibodies so that at least one can bind to the antigen of interest.
+Though the general structure of the mtd protein is constant and very stable, its active ends are extremely variable. Each phage produces a slightly different structural variant of the protein, with the hope that at least one phage can successfully bind to the host and infect it- akin to the way our immune system produces numerous variants of antibodies so that at least one can bind to the antigen of interest.
-The mtd gene, extracted from the BPP-1 phage genome, has the capability of generating a diverse library of stable proteins. Our team hopes to develop a fully in-vitro system to express and select for mtd variants of interest.
+[you should put some background info here about how people have done phage display with this system in the past] Our team hopes to develop a fully in-vitro system to express and select for mtd variants of interest.
 We plan on first generating a diverse DNA library of the Mtd gene, then using mRNA-display to detect and screen for the variants of the protein that can bind to a selected target. In our case, the target will be the surface of E. Coli bacterial cells.