Difference between revisions of "Team:Freiburg/Improve"
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<h1 align="center">Improve</h1> | <h1 align="center">Improve</h1> | ||
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| + | <h2>Hypoxia</h2> | ||
| + | <p>As an improved BioBrick we sent in <i>HRE-pTal</i>. Currently there is no BioBrick containing a minimal promoter in addition to the <i>HRE</i>, but only <i>HRE</i> itself (BBa_K1456004) as well as a combination of <i>CMV-HRE-TATAbox</i> (BBa K1720002). </p> | ||
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| + | <p>Since BBa K1720002 contains a <i>CMV</i> promoter, sensitive oxygen dependent gene expression is not possible. Having a high basal expression downstream of the <i>CMV</i> promoter, BBa K1720002 is not feasible for many projects. This problem is fixed in BBa_K2295003 by introducing the minimal promoter <i>pTal</i> downstream of the <i>HRE</i>, instead of a <i>CMV</i> promoter upstream of the <i>HRE</i>. However, to remove an EcoRI restriction side, the <i>pTal</i> promoter was slightly modified by changing one base pair (detailed description link = link zu BBa_K2295003).</p> | ||
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| + | <p>BBa_K1456004 doesn’t contain a promoter at all. Hence it is not ready to use and a lot of further cloning is involved, before oxygen dependent gene expression can be achieved.<br> | ||
| + | As already mentioned in the “BioBrick Basic Part” (link to https://2017.igem.org/Team:Freiburg/Basic_Part) section, the introduction of multiple enhancer elements allows highly specific oxygen dependent expression in different cell types. Tuning the amount of enhancer elements, downstream transcription can be regulated. <br> | ||
| + | By having a BglII restriction site in between the HRE and the pTal promoter, we offer an easy cloning strategy to change the amount of multiple enhancer elements. Using compatible end cloning, future iGEM teams will profit a lot from the improvement of BBa K1720002 and BBa_K1456004. </p> | ||
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Revision as of 17:05, 28 October 2017
Improve
Hypoxia
As an improved BioBrick we sent in HRE-pTal. Currently there is no BioBrick containing a minimal promoter in addition to the HRE, but only HRE itself (BBa_K1456004) as well as a combination of CMV-HRE-TATAbox (BBa K1720002).
Since BBa K1720002 contains a CMV promoter, sensitive oxygen dependent gene expression is not possible. Having a high basal expression downstream of the CMV promoter, BBa K1720002 is not feasible for many projects. This problem is fixed in BBa_K2295003 by introducing the minimal promoter pTal downstream of the HRE, instead of a CMV promoter upstream of the HRE. However, to remove an EcoRI restriction side, the pTal promoter was slightly modified by changing one base pair (detailed description link = link zu BBa_K2295003).
BBa_K1456004 doesn’t contain a promoter at all. Hence it is not ready to use and a lot of further cloning is involved, before oxygen dependent gene expression can be achieved.
As already mentioned in the “BioBrick Basic Part” (link to https://2017.igem.org/Team:Freiburg/Basic_Part) section, the introduction of multiple enhancer elements allows highly specific oxygen dependent expression in different cell types. Tuning the amount of enhancer elements, downstream transcription can be regulated.
By having a BglII restriction site in between the HRE and the pTal promoter, we offer an easy cloning strategy to change the amount of multiple enhancer elements. Using compatible end cloning, future iGEM teams will profit a lot from the improvement of BBa K1720002 and BBa_K1456004.
