Team:IvyTech SouthBend IN
From 2014.igem.org
Jcampbell188 (Talk | contribs) |
Jcampbell188 (Talk | contribs) |
||
Line 87: | Line 87: | ||
<img src="https://static.igem.org/mediawiki/2014/0/0a/DwikitemplateB_dnastrand.png"> | <img src="https://static.igem.org/mediawiki/2014/0/0a/DwikitemplateB_dnastrand.png"> | ||
<h3>What is your project about?</h3> | <h3>What is your project about?</h3> | ||
- | <p><iframe width="560" height="315" src="//www.youtube.com/embed/6AikSQlZ_nw?list=UUa1eOmuSRxiWnQCV6CElxbQ" frameborder="0" allowfullscreen></iframe> | + | <p><iframe width="560" height="315" src="//www.youtube.com/embed/6AikSQlZ_nw?list=UUa1eOmuSRxiWnQCV6CElxbQ" frameborder="0" allowfullscreen></iframe></p> |
- | The American Public Health Assay (APHA) for human waste contaminated water is hampered by a dependency on culturing coliform bacteria with a turn-around time exceeding one day. An assay for coliphage detection, as surrogate for coliforms, has been developed which employs the release of the host indicator cell β-galactosidase enzyme (β-gal) activity by lytic coliphage and the conversion of a colorimetric substrate. While significantly shortening the APHA time to 3 hrs., this assay requires a laboratory equipment to separate phage-induced released β-gal enzyme from intact cells before addition of the substrate. To optimize this assay we have taken advantage of the α-complementation feature of β-gal. We are engineering cell lines containing either the gene fragments of LacZα or LacZΩ of β-gal on separate plasmids. We predict upon lysis of a mix population of cells by coliphage in the presence of substrate, the gene products will complement in trans reconstituting the β-gal activity thus eliminating a separation step in the assay. In addition we are testing the introduction of a cassette of coliphage cell lysis genes on both plasmids under the control of a coliphage promoter to accelerate the enzyme fragment release thus further reducing test time. Our goal is to design a fast-acting hand-held coliphage biosensor device that can be used by someone without any special technical expertise. We anticipate our device being useful in rapidly reporting the contamination of recreational waters in the U.S. or assuring potable water in the water sources in the developing World. </p> | + | <p>The American Public Health Assay (APHA) for human waste contaminated water is hampered by a dependency on culturing coliform bacteria with a turn-around time exceeding one day. An assay for coliphage detection, as surrogate for coliforms, has been developed which employs the release of the host indicator cell β-galactosidase enzyme (β-gal) activity by lytic coliphage and the conversion of a colorimetric substrate. While significantly shortening the APHA time to 3 hrs., this assay requires a laboratory equipment to separate phage-induced released β-gal enzyme from intact cells before addition of the substrate. To optimize this assay we have taken advantage of the α-complementation feature of β-gal. We are engineering cell lines containing either the gene fragments of LacZα or LacZΩ of β-gal on separate plasmids. We predict upon lysis of a mix population of cells by coliphage in the presence of substrate, the gene products will complement in trans reconstituting the β-gal activity thus eliminating a separation step in the assay. In addition we are testing the introduction of a cassette of coliphage cell lysis genes on both plasmids under the control of a coliphage promoter to accelerate the enzyme fragment release thus further reducing test time. Our goal is to design a fast-acting hand-held coliphage biosensor device that can be used by someone without any special technical expertise. We anticipate our device being useful in rapidly reporting the contamination of recreational waters in the U.S. or assuring potable water in the water sources in the developing World. </p> |
<p></p> | <p></p> | ||
</td> | </td> |
Revision as of 16:34, 3 October 2014
Team:IvyTech SouthBend IN/Templates/Navigationbar
|
|
|
|
|
|