Virginia Tech/Updates/Phage
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- | < | + | <center><h1><font color="#ff8c00"> Phage Culture Updates: |
+ | </font></h1></center> | ||
+ | <center><h3><font color="#8b0000"> VT iGEM Project 2007: Engineering an Epidemic</font></h3></center> | ||
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- | < | + | <p><h3>Introduction: Bacteriophage Lambda</h3> |
- | + | Lambda phage is a well-known and extensively used vector in molecular biology. Lambda is a bacteria virus that can take two different pathways after entering a cell: | |
+ | <html><ul> | ||
+ | <li><b>The most common pathway is the lytic pathway.</b> Upon entering the cell, the virus commands the | ||
+ | cellular machinery to make new phage particles and, within about 45 minutes, lyses the cell.</li> | ||
+ | <li><b>Alternatively, lambda can enter the lysogenic pathway.</b> The viral genome is incorporated into the cell's chromosomal DNA and remains dormant until the right conditions arise (usually involving cellular stress). </li> | ||
+ | </ul></html></p> | ||
+ | <p>Lambda's decision is controlled by the lambda promoter. If enough cI is produced first, the cell becomes lysogenic. If Cro predominates, the cell goes lytic and bursts.<br> | ||
- | < | + | [[Image:Lambda_Pr.JPG]]</p> |
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- | < | + | <p><h3>Updates</h3></p> |
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- | < | + | <p>'''First Progress Update'''<br> |
- | + | ''July 11, 2007''</p> | |
+ | <p>Since no one in our lab had worked with phage before, being able to create and use the lysates has been a learning experience. We have relied heavily on Tim Larson, a faculty adviser from Biology, to help us work through the issues we've come across. We are currently working with two types of phage: Lambda ATCC, a wild-type phage, and Lambda EYFP, a fluorescent phage. Here is what we've accomplished so far with each:</p> | ||
- | < | + | <p>Lambda ATCC:</p> |
- | + | <html><ul> | |
+ | <li>1) Create a high-titer lysate.</li> | ||
+ | <li>2) Infect plates of all 3 strains of E. coli we are working with: LE392, ATCC C600, and a NEB strain. All 3 form plaques approximately equally. </li> | ||
+ | <li>3) View lysis in liquid cultures.</li> | ||
+ | <li>4) Pending: Examine how liquid cultures behave with different amounts of phage using a microtiter plate reader which takes OD readings over time.</li> | ||
+ | </ul></html> | ||
- | <html>< | + | <p>Lambda EYFP:</p> |
- | < | + | <html><ul> |
+ | <li>1) Induce NM538 lysogens and create a lysate.</li> | ||
+ | <li>2) Infect plates of all 3 strains of bacteria we are working with: LE392, ATCC C600, and a NEB strain. The LE392 plate formed plaques at higher dilutions than did either the C600 or NEB plates, which performed the same.</li> | ||
+ | <li>3) Pending: Understand why the LE392 plate performed differently.</li> | ||
+ | <li>4) Pending: View the fluorescence, if possible, using microscopy.</li> | ||
+ | </ul></html> | ||
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- | < | + | <html><center><A href="https://2007.igem.org/Virginia_Tech/Updates" target=_top><h3><FONT color=#ff8c00>Return to Progress Updates</FONT></h3></A></center></html> |
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Latest revision as of 16:51, 24 July 2007
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