Difference between revisions of "Part:BBa K1078001"

Latest revision as of 18:28, 27 September 2013

Strong reporter device optimized for Pichia pastoris, activated by methanol

This device is composed of three parts. The first part is a modified promoter P_AOX1; it was found in a library where it showed more than 60% stronger activation when compared with the wild-type P_AOX1 (found as BBa_K431007 in the registry of parts), as reported by Hartner FS et al., in Promoter library designed for fine-tuned gene expression inPichia pastoris. Nucleic Acids Res 2008, 36:e76. The second part is a kozak sequence for P. pastoris, and the third is a red fluorescent protein condon optimized for expression in P. pastoris.

This device was created to express a reporter gene, in this case RFP, in the presence of methanol. Its expression is inactivated by ethanol. When combined with the modified Mxr1 (BBa_K1078000), it is not inactivated by ethanol. This is useful for our biosensor design, which aims to detect levels of methanol above 2% in common alcoholic drinks (normally containing 10 to 60% ethanol). This will allow government to make a preliminary high-throughput screening of ethanol drinks tainted will methanol.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K1078001 short</partinfo>
-This device is composed by three parts. The first part is a modified promoter pAOX, it was found in a library where it showed more than 60% stronger activation when compared with the wild-type, as reported by Hartner FS, Ruth C, Langenegger D, Johnson SN, Hyka P, Lin-Cereghino GP, Lin-Cereghino J, Kovar K, Cregg JM, Glieder A: Promoter library designed for fine-tuned gene expression in Pichia pastoris. Nucleic Acids Res 2008, 36:e76. The second part is the kozak sequence for Pichia pastoris, and the third is a Red fluorescent protein condon optimized for expression in Pichia pastoris.
+<html>
-This device was created to express a reporter gene in the presence of ethanol, and is inactivated by ethanol, when combined with the modified Mxr1 (BBa_K1078000), it is not inactivated by ethanol. This is useful for our biosensor design, which aims to detect levels of methanol above 2% in common alcoholic drinks (normal content of 10 to 60% ethanol). This will allow government to make high-throughput screening of ethanol drinks tainted will methanol.
+<p>This device is composed of three parts. The first part is a modified promoter P<sub>AOX1</sub>; it was found in a library where it showed more than 60% stronger activation when compared with the wild-type P<sub>AOX1</sub> (found as <a href="https://parts.igem.org/Part:BBa_K431007">BBa_K431007</a> in the registry of parts), as reported by Hartner FS et al., in <i>Promoter library designed for fine-tuned gene expression inPichia pastoris. Nucleic Acids Res 2008, 36:e76</i>. The second part is a kozak sequence for <i>P. pastoris</i>, and the third is a red fluorescent protein condon optimized for expression in <i>P. pastoris</i>.</p>
+<p>This device was created to express a reporter gene, in this case RFP, in the presence of methanol. Its expression is inactivated by ethanol. When combined with the modified Mxr1 (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1078000">BBa_K1078000</a>), it is not inactivated by ethanol. This is useful for our biosensor design, which aims to detect levels of methanol above 2% in common alcoholic drinks (normally containing 10 to 60% ethanol). This will allow government to make a preliminary high-throughput screening of ethanol drinks tainted will methanol. </p>
-The modified Mxr1 is able to activate the pAOX promoter, in ethanol, glycerol and methanol. This is useful for our biosensor design, which aims to detect levels of methanol above 2% in common alcoholic drinks (normal content of 10 to 60% ethanol). This will allow government to make high-throughput screening of ethanol drinks tainted will methanol.
+</html>
 <!-- Add more about the biology of this part here