Team:Nevada/misc

From 2010.igem.org

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Latest revision as of 22:49, 15 October 2010

ccdB

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The ccdB gene is a known topoisomerase II poison and will kill all available cell lines with the exception of DB3.1 cell lines available from Invitrogen. The gene can be used as a selectable marker by ligating it with the plasmid and structure of interest and transforming it into a cell line susceptible to the toxic effects of ccdB. Colonies that grow on plates should contain the plasmid and gene of interest as ccdB inserted into the plasmid will kill cell cultures.

We would like to thank the following sponsors for their support in helping us make this project possible. Much thanks to the [http://www.cabnr.unr.edu/Students/Majors.aspx Departments of Biochemistry and Biotechnology] and the [http://www.cabnr.unr.edu/Students/Glance.aspx College of Agriculture, Biotechnology and Natural Resources] for their encouragement and support. Thank you [http://www.unr.edu/inbre/ Nevada INBRE] for over $6,000 in support for supplies and registration costs. Thank you to Associated Students of the Univeristy of Nevada for supporting our fund raising efforts. Thank you to [http://www.promega.com/Catalog/CountrySelect.aspx?returnurl=/Default.asp Promega Co.] for free enzyme donations. Thank you to [http://www.invitrogen.com/site/us/en/home.html?cid=covinvggl89100000002336s& Invitrogen Co.] for a discount on our Vector NTI program.

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-== NT Cell Transformations ==
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-<p>The 2010 Nevada iGEM team has three goals for this year’s competition.  First, we are going to test the validity of utilizing Nicotiana tabacum protoplasts (NT cells), plant cells without the cell wall, as a model for the expression of higher plant genes for future iGEM competitions.  This system is useful in the respect that the time it takes to obtain transgenic lines of cells is greatly reduced compared to the time to obtain transgenic plants.  <html>
+== ccdB ==
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 <a href="https://2010.igem.org/Image:IMG_6031.JPG"><img src="https://2010.igem.org/Image:IMG_6031.JPG" class="shadow" style="float:left;width:170px;margin:10px"></a>
-</html>These cells can therefore be utilized as a quick proof-of-concept test model before moving synthetic constructs into plants of interest.  We also aim to produce an iGEM-compatible plant-specific plasmid, several stress-inducible plant promoters, reporter genes containing Kozak sequences (ribosome binding sites) and terminators that conform to BioBrick standards.  Lastly, we hope to measure the induction of these stress promoters in real-time by performing a fluorometry assay in which stress will be applied to NT cells and fluorescent output by a reporter (GFP) will be measured to detail induction in real time.  This method has a distinct advantage over microarrays since microarrays are only one ‘snapshot’ in time.</p>
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+The ccdB gene is a known topoisomerase II poison and will kill all available cell lines with the exception of DB3.1 cell lines available from Invitrogen.  The gene can be used as a selectable marker by ligating it with the plasmid and structure of interest and transforming it into a cell line susceptible to the toxic effects of ccdB.  Colonies that grow on plates should contain the plasmid and gene of interest as ccdB inserted into the plasmid will kill cell cultures.